JP2004512264A - Treatment of human papillomavirus - Google Patents
Treatment of human papillomavirus Download PDFInfo
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- JP2004512264A JP2004512264A JP2002505023A JP2002505023A JP2004512264A JP 2004512264 A JP2004512264 A JP 2004512264A JP 2002505023 A JP2002505023 A JP 2002505023A JP 2002505023 A JP2002505023 A JP 2002505023A JP 2004512264 A JP2004512264 A JP 2004512264A
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Abstract
（１）熱ショックタンパク質、またはその免疫刺激性断片、および（２）ヒト乳頭腫ウイルスのタンパク質、またはその抗原性断片を含む組成物を、対象に投与することで、対象の疣贅を治療する方法を開示する。また第１の型のヒト乳頭腫ウイルスに感染した対象、または感染が疑われる対象におけるヒト乳頭腫ウイルス感染を、（１）熱ショックタンパク質、またはその抗原性断片、および（２）第２の型のヒト乳頭腫ウイルスのタンパク質、またはその抗原性断片を含む組成物を対象に投与することにより、治療する方法であって、第１の型および第２の型は異なる方法も開示する。Treating a subject with a composition comprising (1) a heat shock protein, or an immunostimulatory fragment thereof, and (2) a composition of a human papillomavirus protein, or an antigenic fragment thereof, to the subject. A method is disclosed. In addition, human papillomavirus infection in a subject infected or suspected of being infected with a first type of human papillomavirus, comprises: (1) a heat shock protein, or an antigenic fragment thereof, and (2) a second type. Also disclosed are methods of treating by administering to a subject a composition comprising a human papillomavirus protein, or an antigenic fragment thereof, wherein the first type and the second type are different.
Description
【０００１】
発明の分野
本発明は、ヒト乳頭腫ウイルス感染の治療法に関する。
【０００２】
発明の背景
ヒト乳頭腫ウイルス（ＨＰＶ）感染は広くみられる。ＨＰＶは性行為によって伝達され、性的に活発な成人の２０〜８０％が感染していると推定されている。感染の大半は無症候性であるが、感染すると性器疣贅の発生（発生率は成人の約１〜５％）、およびの陰部の癌の発生に至る場合がある。別のタイプの癌である子宮頚癌はＨＰＶに強く関連している（Ｆｒａｚｅｒ、Ｇｅｎｉｔｏｕｒｉｎ． Ｍｅｄ． ７２：３９８〜４０３、１９９６）。ＨＰＶの６型、１１型、１６型、１８型、３１型、および３３型は、癌の危険性の上昇と関連する場合があり、１６型および／または１８型が子宮頚癌の９０％より多くで検出されている（ｖａｎ Ｄｒｉｅｌら、Ａｎｎ． Ｍｅｄ． ２８：４７１〜４７７、１９９６）。６型および１１型も肛門性器の疣贅と関連している。乳頭腫ウイルスと関連病変に関する総説は、シャー（Ｓｈａｈ）ら、「ウイルス学（Ｖｉｒｏｌｏｇｙ）」第３版の「第６６章：乳頭腫ウイルス（Ｃｈａｐｔｅｒ ６６：Ｐａｐｉｌｌｏｍａｖｉｒｕｓｅｓ）」（Ｆｉｅｌｄｓら編、Ｒａｖｅｎ Ｐｒｅｓｓ、Ｐｈｉｌａｄｅｌｐｈｉａ、ｐｐ ２０７７〜２１０９、１９９６）、およびハウゼン（ｚｕｒ Ｈａｕｓｅｎ）、Ｊ． Ｎａｔｌ． Ｃａｎｃｅｒ Ｉｎｓｔ． ９２：６９０〜６９８、２０００を参照されたい。
【０００３】
現在、免疫系を標的とすることで疣贅または上述の疾患を治療または予防する安全で有効な方法はない。このような治療法を開発する取り組みは、いくつかの理由で進んでいない。一つの理由は、１種類のＨＰＶ型に由来する抗原は、限られた型特異的な免疫応答しか生じないという定説の存在である。したがって、多種多様なＨＰＶ型に由来する抗原を含むカクテルが、広範囲に有効なＨＰＶ療法に必要であることが示唆されている（Ｃａｉｎｅら、Ｓｃｉｅｎｃｅ ２８８：１７５３、２０００）。
【０００４】
発明の概要
本発明は、１種類のＨＰＶ型に由来するタンパク質を含む融合タンパク質が、別のＨＰＶ型の感染で引き起される疾患または症状の治療に使用可能であるという発見に部分的に基づく。例えば細菌の熱ショックタンパク質（ｈｓｐ）と融合させたＨＰＶ １６型の抗原は、１６型以外のＨＰＶ型（例えばＨＰＶ ６型および１１型）を原因とするヒトの肛門性器疣贅の治療に有効であった。この結果は以下の２つの点を支持する：（１）疣贅がＨＰＶタンパク質で治療可能であること、また（２）ＨＰＶに対する治療薬が、広域の有効性を目的として、異なるＨＰＶ型に由来するタンパク質抗原を含む必要がないこと。
【０００５】
したがって本発明は、（１）ｈｓｐ、またはその免疫刺激性断片、および（２）ＨＰＶタンパク質（例えばＨＰＶ １６型のＥ７タンパク質などの抗原タンパク質）、またはその抗原性断片を含む組成物を対象に投与することで、対象の疣贅を治療する方法を特徴とする。このような組成物は、それぞれ「成分（１）」と「成分（２）」と本明細書では呼ぶことにする。ｈｓｐ （またはその免疫刺激性断片）およびＨＰＶタンパク質（またはその抗原性断片）は、同じ調製物中に単純に混合するか、または化学的接合または融合で密接に結合させることができる（すなわち、本明細書に記載された成分を含む融合タンパク質、またはそれをコードする核酸を投与することができる）。混合、接合、または融合することで、成分（１）と成分（２）を同時に投与することができる。しかし各成分を分けて（例えば連続的に）投与したり、成分（１）なしに成分（２）を投与したりすることができる。上述の方法には、疣贅を有する対象、または疣贅を有することが疑われる対象を同定する段階を含めることができる（対象を治療するという意味において、同定は治療薬剤の投与開始前になされる場合がある）。医師および他の当業者は、このような患者を適切に同定することができる。
【０００６】
本発明の方法は、（既に疣贅を有する対象ではなく）、疣贅予防を望む対象、または疣贅予防で利益を得る対象が同定される場合に、疣贅の予防に用いることもできる。
【０００７】
本発明はまた、（１）ｈｓｐ、またはその免疫刺激性断片、および（２）第２の型（例えば１６型）のＨＰＶのタンパク質、またはその抗原性断片を含む組成物を対象に投与することで、第１の型（例えば５型、６型、１１型、１８型、３１型、３３型、３５型、４５型、５４型、６０型、または７０型）のＨＰＶ感染が原因の疾患または症状を有する対象を治療する方法を特徴とする。「第１の型」のＨＰＶ、および「第２の型」のＨＰＶは互いに異なり、２つの異なるＨＰＶの型である。ｈｓｐ （またはその免疫刺激性断片）、およびＨＰＶタンパク質（またはその抗原性断片）は、同じ調製物中に単純に混合するか、または化学的な接合または融合で、より密接に結合させることができる（すなわち、本明細書に記載された成分を有する融合タンパク質、またはそれをコードする核酸分子を投与することができる）。混合、接合、または融合することで、成分（１）と成分（２）を同時に投与することができる。一方、各成分を分けて（例えば連続的に）投与したり、成分（１）なしに成分（２）を投与したりすることができる。さらに、このような方法にはＨＰＶに感染した対象、または感染（または関連する疾患または症状）が疑われる対象を同定する段階を含めることができる。
【０００８】
第１のＨＰＶ型の感染者に、第２の型のＨＰＶを含む組成物を投与することで、上記の方法を、ＨＰＶ感染を分類する前に、それが顕在化する前に、または症状が現れる前に実施することができる（すなわち、対象が感染している、または感染するおそれのある特定のＨＰＶ型を、治療または予防の開始前に知る必要はない）。この方法が予防的な場合、方法にはＨＰＶ感染の予防を望む対象、または予防による利益を得る対象を同定する段階を含めることができる。
【０００９】
本明細書に記載された組成物は、（例えば疣贅の大きさを小さくすること、もしくは疣贅の形状を変えることで、または疣贅関連症状（例えば足底疣贅に伴うことの多い疼痛）を緩和させることで）疣贅の治療に十分な量を投与することが可能であり、対象に複数の疣贅がみられる場合は、疣贅の個数を減らす段階を治療に含めることができる。本明細書に記載された組成物は同様に、ＨＰＶ感染が原因の疾患、またはＨＰＶ感染に関連する疾患（例えば子宮頚癌や肛門癌などの癌、または、例えば子宮頚部異形成もしくは肛門異形成など他の症状）の治療に十分な量を投与することができる。疣贅は上述のように個別に言及されるが、疣贅はＨＰＶに起因する状態、またはＨＰＶに関連した状態を含む場合もある。医師および他の当業者は、疣贅または疾患もしくは状態に関連する望ましくない生理学的作用が減少する場合に、疣贅もしくはＨＰＶが関連する疾患または症状に有効な「治療法」を理解すると思われる。疣贅の臨床症状および生理的症状、ならびにＨＰＶ感染に関連した疾患または症状の臨床症状および生理的症状については、例えばファウチ（Ｆａｕｃｉ）ら、「ハリソンの内科学の原理（Ｈａｒｒｉｓｏｎ’ｓ Ｐｒｉｎｃｉｐｌｅｓ ｏｆ Ｉｎｔｅｒｎａｌ Ｍｅｄｉｃｉｎｅ）」、第１４版、ＭｃＧｒａｗ−Ｈｉｌｌ Ｐｒｅｓｓ、Ｎｅｗ Ｙｏｒｋ、ｐｐ ３０２〜３０３および１０９８〜１１００、１９９８で検討されている。
【００１０】
本明細書に記載された方法で利益を得る「対象」は、乳頭腫ウイルスに感染可能な動物（例えばヒトや家畜（例えば、ウシ、ウマ、ブタ、ヒツジ、およびヤギ）などの哺乳類、ならびに家畜動物（例えばネコやイヌ）である。疣贅は、対象の陰部、皮膚、または内臓（再発性呼吸器乳頭腫症（ＲＲＰ；若年性喉頭乳頭腫症（ＪＬＰ）または成人発症ＲＲＰとも呼ばれる）における声帯に現れる疣贅など）に生じる場合がある。
【００１１】
本発明は、本明細書に記載された方法による、疣贅またはＨＰＶ感染に関連した（もしくはＨＰＶ感染を原因とする）疾患もしくは状態を有する対象の治療に用いる（タンパク質、タンパク質抱合体（ｃｏｎｊｕｇａｔｅ）、もしくは融合タンパク質、またはこれらをコードする核酸分子を含む）本明細書に記載された１種または複数の組成物の用途をさらに含む。本発明はまた本明細書に記載された方法による、疣贅またはＨＰＶ感染に関連する（または感染を原因とする）疾患もしくは状態を有する対象治療用の薬物製造における１種または複数のこのような組成物の用途も含む。
【００１２】
タンパク質（例えばＨＰＶタンパク質）の「抗原性断片」は、本明細書に記載された方法で投与されたときに、タンパク質に対して断片特異的またはその断片が由来するタンパク質特異的な免疫応答を対象に生じるタンパク質の任意の部分である。免疫応答は液性反応または細胞性反応のいずれの場合もありうる。例えば抗原性断片は、後述するＨＬＡクラスＩペプチド抗原の場合がある。当業者であれば、本発明の文脈で望ましい免疫応答が、完全なタンパク質およびタンパク質断片で生じるだけでなく、変異型タンパク質（例えば１か所もしくは複数の箇所におけるアミノ酸配列の付加、置換（例えば保存的アミノ酸置換）、または欠失を含むタンパク質）でも生じることがありうることを理解すると思われる。変異型ＨＰＶ抗原の作製、および本発明の文脈における作用の検討は容易である。
【００１３】
タンパク質（例えばｈｓｐ）の「免疫刺激性断片」は、本発明にしたがって投与されたときに、抗原による免疫応答を促進するタンパク質の任意の部分である。例えばＨＰＶタンパク質に対する免疫応答が、ＨＰＶタンパク質がｈｓｐの断片とともに（例えば融合した状態で）投与されたときに促進される場合、そのような断片はｈｓｐの免疫刺激性断片である。当業者であれば、このような免疫応答が、変異型ｈｓｐ （例えばアミノ酸配列中に１か所または複数の箇所における付加、置換（例えば保存的アミノ酸置換）、または欠失を含むｈｓｐ）でも促進されることを理解すると思われる。変異型ｈｓｐは、その作製およびＨＰＶ抗原に対する免疫応答促進能力の検討が容易である。
【００１４】
本発明の方法は以下の段階の効率のよい手段を提供する：（１）疣贅を治療または予防する段階、および（２）他の型のＨＰＶを含む組成物による（すなわち組成物を使用する）、あるＨＰＶ型の感染を原因とする（または感染に関連する）疾患もしくは状態を治療または予防する段階。したがって、あるＨＰＶ型のＨＰＶ抗原を含む組成物を、感染した（または感染する可能性のある）ＨＰＶ型にかかわらず、大半ではなくても多くの対象で使用することができる。ＨＰＶ組成物が、このような環境（交差反応を必要とする環境）で有効性を示すことは驚くべきことである。一つの型のＨＰＶ抗原は、別の型に対して有効な免疫応答を生じないとこれまで考えられきた。本発明の他の特性または利点は、以下の詳細な説明および特許請求の範囲からも明らかになる。
【００１５】
詳細な説明
本発明は、ｈｓｐおよびＨＰＶタンパク質（例えばタンパク質抗原）を含む、広範囲に有効なＨＰＶを基礎とした治療薬に関する。特定の機構を介して作用を生じるＨＰＶに基づく治療法に本発明を制限することなく、このような薬剤は、疣贅およびＨＰＶ感染に関連した他の状態（例えば異形成）、および疾患（例えば癌）を改善する免疫応答を生じると考えられている。例えば本発明の組成物は複数種のＨＰＶ型に由来するＨＰＶタンパク質を含む場合があるが、一つの型にのみ由来するＨＰＶタンパク質を含む場合がある。また、一つのＨＰＶ型に由来するＨＰＶタンパク質を含む組成物は、他の型のＨＰＶ感染で生じる疣贅またはＨＰＶに関連する他の疾患または症状の治療または予防に有用である。本発明に関連した用途に適したさまざまな材料および手順を以下に述べる。
【００１６】
融合タンパク質の調製
ｈｓｐおよびＨＰＶタンパク質をコードする核酸配列は当業者に知られており、また利用されている。したがって、本発明の方法に有用な融合ポリペプチドをコードする核酸構築物は、日常的な方法で容易に調製することができる（同様に、このような核酸分子はｈｓｐおよびＨＰＶタンパク質の作製に個別に使用することができるほか、個々のｈｓｐおよびＨＰＶタンパク質を後に（例えば単純に混合して）物理的に結合させたり、化学的接合（後述）で、またはジスルフィド結合を介して連結させたりすることができる）。抗原（例えばＨＰＶ抗原）に選択的に融合させたｈｓｐをコードする核酸配列の例は、国際公開公報第８９／１２４５５号、第９４／２９４５９号、第９８／２３７３５号、および第９９／０７８６０号、ならびに本明細書で引用した参考文献に記載されている。融合タンパク質を含むタンパク質を発現させて精製する方法を以下に述べる。
【００１７】
タンパク質抱合体の調製
成分（１）および成分（２）は、各ｈｓｐと各ＨＰＶ抗原との翻訳後結合で連結させることもできる。２つのタンパク質（またはそれらの断片）を化学的に結合する方法は当技術分野で周知である（例えばＨｅｒｍａｎｓｏｎ、「生物抱合体技術（Ｂｉｏｃｏｎｊｕｇａｔｅ Ｔｅｃｈｎｉｑｕｅｓ）」、Ａｃａｄｅｍｉｃ Ｐｒｅｓｓ、Ｓａｎ Ｄｉｅｇｏ、ＣＡ、１９９６；Ｌｕｓｓｏｗら、Ｅｕｒ． Ｊ． Ｉｍｍｕｎ． ２１：２２９７〜２３０２、１９９１；およびＢａｒｒｉｏｓら、Ｅｕｒ． Ｊ． Ｉｍｍｕｎ． ２２：１３６５〜１３７２、１９９２に記載された手法を参照）。抱合体は、グルタルアルデヒド（結果として生じる抱合体の一部となる）などの架橋剤を用いる方法で、または成分（１）と成分（２）をジスルフィド結合で連結する方法で調製することができる。またｈｓｐ中に天然に存在するシステイン残基、もしくは組換え的に挿入したシステイン残基、またはその両方を用いて分子間ジスルフィド結合の形成を促すことができる。ＨＰＶ抗原と非共有結合的に結合するｈｓｐ、またはその免疫刺激性断片（成分（１））を含む組成物は、米国特許第
【００１８】
投与組成物の最終形状にかかわらず、成分（１）および成分（２）は以下に挙げる分子を含む場合がある。
【００１９】
ＨＰＶ タンパク質抗原
任意のＨＰＶ抗原が、本発明の組成物（例えば本明細書に記載された混合物、抱合体、および融合タンパク質）における用途に適している。しかし、認識可能なエピトープをＨＰＶ感染細胞の表面に発現するＨＰＶ抗原は特に有用であるべきである。ＨＰＶは６個または７個の非構造タンパク質、および２個の構造タンパク質を発現し、これらはいずれも本明細書に記載された免疫学的な予防法または免疫学的な治療法における標的として作用する。
【００２０】
ウイルスのカプシドタンパク質Ｌ１およびＬ２は後期構造タンパク質である。Ｌ１は主要カプシドタンパク質であり、そのアミノ酸配列はさまざまなＨＰＶ型でよく保存されている。また７種の初期非構造タンパク質が存在する。タンパク質Ｅ１、Ｅ２、およびＥ４はウイルスの複製に重要な役割を果たす。タンパク質Ｅ４はウイルスの成熟にも役割を果たす。Ｅ５の果たす役割はよくわかっていない。タンパク質Ｅ６およびＥ７は、ウイルスの複製、ならびに宿主細胞の不死化および形質転換に不可欠な癌タンパク質である。
【００２１】
Ｈｓｐ
多様な生物種からさまざまなｈｓｐが単離され、クローン化され、また特性解析が行われている（Ｍｉｚｚｅｎ、Ｂｉｏｔｈｅｒａｐｙ １０：１７３〜１８９、１９９８；本明細書で用いられるように「熱ショックタンパク質」という用語、またはその省略形のｈｓｐは同義であり、「ストレスタンパク質」と呼ばれるタンパク質を含む）。免疫系を刺激するｈｓｐ、またはその免疫刺激性断片は、本明細書に記載された組成物（例えば融合ポリペプチドの一部）における用途に適している。ｈｓｐ７０、ｈｓｐ６０、ｈｓｐ２０〜３０、およびｈｓｐ１０は、結核菌（Ｍｙｃｏｂａｃｔｅｒｉｕｍ ｔｕｂｅｒｃｕｌｏｓｉｓ）およびライ菌（Ｍｙｃｏｂａｃｔｅｒｉｕｍ ｌｅｐｒａｅ）による感染に対する宿主免疫応答によって認識される、主要な決定因子群である。またウシ型結核菌の一つの株であるカルメット−ゲラン桿菌（Ｂａｃｉｌｌｅ Ｃａｌｍｅｔｔｅ Ｇｕｅｒｉｎ；ＢＣＧ）のｈｓｐ６５は、実施例で説明されるように有効な免疫刺激剤であることがわかっている。
【００２２】
ｈｓｐ遺伝子およびｈｓｐのファミリーは、成分（１）について説明されるように使用可能であり、当技術分野で周知である。このファミリーには例えばＨｓｐ１００〜２００、Ｈｓｐ１００、Ｈｓｐ９０、Ｌｏｎ、Ｈｓｐ７０、Ｈｓｐ６０、ＴＦ５５、Ｈｓｐ４０、ＦＫＢＰ、シクロフィリン、Ｈｓｐ２０〜３０、ＣｌｐＰ、ＧｒｐＥ、Ｈｓｐ１０、ユビキチン、カルネキシン、およびタンパク質ジスルフィドイソメラーゼなどが含まれる。これについては例えばマカリオ（Ｍａｃａｒｉｏ）、Ｃｏｌｄ Ｓｐｒｉｎｇ Ｈａｒｂｏｒ Ｌａｂｏｒａｔｏｒｙ Ｒｅｓ． ２５：５９〜７０、１９９５；パーセル（Ｐａｒｓｅｌｌ）ら、Ｒｅｖ． Ｇｅｎｅｔ． ２７：４３７〜４９６、１９９３；および米国特許第５，２３２，８３３号を参照されたい。ｈｓｐは哺乳類、細菌、またはマイコバクテリアのｈｓｐの場合があるがこれらに限定されない。
【００２３】
Ｇｒｐ１７０ （グルコース調節型タンパク質）はｈｓｐ１００〜２００ファミリーのｈｓｐの一例である。Ｇｒｐ１７０は小胞体の内腔、前ゴルジ体に存在し、免疫グロブリンの折りたたみおよび集合に重要な役割を果たす場合がある。
【００２４】
ｈｓｐ１００ ファミリーのｈｓｐの例には哺乳類のＨｓｐ１１０、酵母のＨｓｐ１０４、ならびに大腸菌のｈｓｐ （ＣｌｐＡ、ＣｌｐＢ、ＣｌｐＣ、ＣｌｐＸ、およびＣｌｐＹ）などがある。
【００２５】
ｈｓｐ９０ファミリーの例には大腸菌のＨｔｐＧ、酵母のＨｓｐ８３およびＨｓｃ８３、ならびにヒトのＨｓｐ９０α、Ｈｓｐ９０β、およびＧｒｐ９４などがある。Ｈｓｐ９０は、情報伝達機構に重要な役割を果たすステロイドホルモン受容体（例えばグルココルチコイド、エストロゲン、プロゲステロン、およびテストステロンの受容体）、転写因子、ならびにタンパク質キナーゼなどの通常は細胞調節分子であるタンパク質群に結合する。Ｈｓｐ９０タンパク質はまた、他のストレスタンパク質を含む大型で量の多いタンパク質複合体の形成にも関与する。
【００２６】
Ｌｏｎは、大腸菌の非天然タンパク質を分解する四量体のＡＴＰ依存性プロテアーゼである。
【００２７】
ｈｓｐ７０ファミリーのｈｓｐの例には哺乳類細胞のＨｓｐ７２およびＨｓｃ７３、細菌またはマイコバクテリア（ライ菌、結核菌、およびウシ型結核菌（カルメット−ゲラン桿菌など；本明細書ではｈｓｐ７１と記載）のＤｎａＫ、大腸菌、酵母、および他の原核生物のＤｎａＫ、ならびにＢｉＰおよびＧｒｐ７８などがある。ｈｓｐ７０はＡＴＰ、ならびに折りたたまれていないポリペプチドおよびペプチドに特異的に結合することができる。ｈｓｐ７０はタンパク質の折りたたみおよび解きほぐし、ならびにタンパク質複合体の集合および分解に関与する。
【００２８】
Ｈｓｐ６０ファミリーのｈｓｐの例にはマイコバクテリアのＨｓｐ６５がある。細菌のＨｓｐ６０はＧｒｏＥＬとしても広く知られている。Ｈｓｐ６０は大型のホモオリゴマー複合体を形成し、タンパク質の折りたたみに重要な役割を果たすと考えられている。Ｈｓｐ６０の相同物は真核生物のミトコンドリアおよび葉緑体中に存在する。
【００２９】
ＴＦ５５ファミリーのｈｓｐの例にはＴｃｐ１、ＴＲｉＣ、およびサーモソーム（ｔｈｅｒｍｏｓｏｍｅ）などがある。これらのタンパク質は通常、真核生物および一部の古細菌の細胞質に認められ、タンパク質の折りたたみを促進する多員環を形成する。これらはまたＨｓｐ６０と弱い相同性を有する。
【００３０】
Ｈｓｐ４０ファミリーのｈｓｐの例には大腸菌およびマイコバクテリアなどの原核生物のＤｎａＪ、ならびにＨＳＪ１、ＨＤＪ１、およびＨｓｐ４０などがある。Ｈｓｐ４０は、タンパク質の折りたたみ、熱耐性、およびＤＮＡの複製などの細胞活性における分子シャペロンとして作用する。
【００３１】
ＦＫＢＰの例にはＦＫＢＰ１２、ＦＫＢＰ１３、ＦＫＢＰ２５、ＦＫＢＰ５９、Ｆｐｒ１、およびＮｅｐ１などがある。これらのタンパク質は通常、ペプチジル−プロピルイソメラーゼ活性を有し、ＦＫ５０６やラパマイシンなどの免疫抑制剤と相互作用する。これらのタンパク質は通常、細胞質および小胞体に存在する。
【００３２】
シクロフィリンの例には、シクロフィリンＡ、Ｂ、およびＣなどがある。これらのタンパク質はペプチジル−プロリルイソメラーゼ活性を有し、免疫抑制剤シクロスポリンＡと相互作用する。
【００３３】
Ｈｓｐ２０〜３０は低分子量Ｈｓｐ （ｓｍａｌｌ Ｈｓｐ）とも呼ばれる。Ｈｓｐ２０〜３０は通常、大型のホモオリゴマー複合体中に存在し、ヘテロオリゴマー複合体中にみられることもある。生物体または細胞の種類によっては、複数の異なる種類の低分子量ｈｓｐを発現することがある。Ｈｓｐ２０〜３０は細胞骨格構造と相互作用し、アクチンの重合／脱重合を調節する役割を果たすことがある。Ｈｓｐ２０〜３０は、ストレスを受けたり、休止細胞が成長因子に曝されたりすると速やかにリン酸化される。Ｈｓｐ２０〜３０の相同物にはα−クリスタリンなどがある。
【００３４】
ＣｌｐＰは、異常なタンパク質の分解に関与する大腸菌由来のプロテアーゼである。ＣｌｐＰの相同物は葉緑体に存在する。ＣｌｐＰはＣｌｐＡとヘテロオリゴマー複合体を形成する。
【００３５】
ＧｒｐＥは、ストレス損傷を受けたタンパク質の救済、ならびに損傷タンパク質の分解に関与する約２０ ｋＤａの大腸菌タンパク質であり、ＧｒｐＥは大腸菌におけるストレス遺伝子の発現調節に重要な役割を果たす。
【００３６】
Ｈｓｐ１０の例にはＧｒｏＥＳやＣｐｎ１０などがある。Ｈｓｐ１０は大腸菌、ならびに真核細胞のミトコンドリアおよび葉緑体にみられる。Ｈｓｐ１０は、Ｈｓｐ６０オリゴマーと結合する七員環を形成する。Ｈｓｐ１０はタンパク質の折りたたみにも関与する。
【００３７】
ユビキチンは、ＡＴＰ依存性細胞質プロテアーゼによるタンパク質のタンパク質分解性の除去を調整するタンパク質に結合することがわかっている。
【００３８】
本発明で有用なストレスタンパク質は、腸内細菌（例えば大腸菌）、マイコバクテリア（特に、ライ菌、結核菌、マイコバクテリウム・バッカエ（Ｍ． ｖａｃｃａｅ）、恥垢菌、およびウシ型結核菌）、酵母、ショウジョウバエ、脊椎動物（例えば鳥類、またはげっ歯類もしくはヒトを含む霊長類などの哺乳類）から得られる。
【００３９】
タンパク質の発現および精製
タンパク質は組換え的に作製することができる。具体的には、個別投与、併用投与、または結合後の投与が可能なｈｓｐ （またはその断片）およびＨＰＶ抗原（またはその断片）、ならびに成分（１）および成分（２）を含む融合タンパク質を細菌、酵母、植物もしくは植物細胞、または動物もしくは動物細胞で組換え的に作製することができる。例えばｈｓｐ、ＨＰＶ抗原、およびこれらを含む融合タンパク質を、宿主細胞を対象に、適切な発現媒体上の核酸配列を用いる形質転換法（トランスフェクション、形質導入、または感染）で作製することができる。適切な発現媒体には、プラスミド、ウイルス粒子、およびファージなどがある。昆虫細胞にはバキュロウイルス発現ベクターなどが適している。発現媒体の全体または一部を宿主細胞ゲノムに組み込ませることができる。状況によっては、誘導型発現ベクター、例えばＬＡＣＳＷＩＴＣＨ （登録商標）の「誘導発現システム（Ｉｎｄｕｃｉｂｌｅ Ｅｘｐｒｅｓｓｉｏｎ Ｓｙｓｔｅｍ）」（Ｓｔｒａｔａｇｅｎｅ；Ｌａ Ｊｏｌｌａ、ＣＡ）を使用することが望ましい。
【００４０】
分子生物学分野の当業者であれば、任意のさまざまな発現系を用いることで、本明細書に記載された方法に有用な組換えタンパク質（例えば融合タンパク質）を提供できることを理解すると思われる。使用する正確な宿主細胞およびベクターは本発明に重要ではない。
【００４１】
上述したように、成分（１）、成分（２）、およびこれらを含む融合タンパク質を植物細胞で作製することができる。植物細胞には、ウイルス発現ベクター（例えばカリフラワーモザイクウイルスおよびタバコモザイクウイルス）、ならびにプラスミド発現ベクター（例えばＴｉプラスミド）が適している。このような細胞は、さまざまなな供給先から入手できる（例えば米国菌培養収集所（ＡＴＣＣ）、Ｍａｎａｓｓａｓ、ＶＡ；例えばＡｕｓｕｂｅｌら、「分子生物学の最新プロトコール（Ｃｕｒｒｅｎｔ Ｐｒｏｔｏｃｏｌｓ ｉｎ Ｍｏｌｅｃｕｌａｒ Ｂｉｏｌｏｇｙ）」、Ｊｏｈｎ Ｗｉｌｅｙ ＆ Ｓｏｎｓ、Ｎｅｗ Ｙｏｒｋ、１９９４も参照）。形質転換の方法および発現媒体の選択は選択される宿主系によって変わる。形質転換の方法については例えばオースベル（Ａｕｓｕｂｅｌ）の前掲書に記載されている。発現媒体は、例えばパウエルズ（Ｐｏｕｗｅｌｓ）ら、「クローニングベクター：実験マニュアル（Ｃｌｏｎｉｎｇ Ｖｅｃｔｏｒｓ：Ａ Ｌａｂｏｒａｔｏｒｙ Ｍａｎｕａｌ）」、１９８５、Ｓｕｐｐ． １９８７に記載されたものから選択することができる。
【００４２】
発現媒体をもつ宿主細胞は、選択遺伝子の活性化または抑制、形質転換体の選択、または選択遺伝子の増幅に対して必要に応じて適合させた従来の栄養培地中で培養することができる。
【００４３】
適切であれば、または有益であれば、融合タンパク質をコードする核酸に、融合タンパク質を細胞培養物中に分泌させて同培養物中からタンパク質の分離を促すためのシグナル配列を含めることができる。特定の開始シグナルが、挿入核酸配列の効率のよい翻訳に必要な場合もある。このようなシグナルにはＡＴＧ開始コドンおよび隣接配列が含まれる。場合によっては、できればＡＴＧ開始コドンを含む外因性の翻訳制御シグナルを提供しなくてはならない。また開始コドンは、挿入全体を確実に翻訳させるために、所望のコード配列の読み枠と相があっていなければならない。このような外因性翻訳制御シグナルおよび開始コドンは、さまざまな起源のものを天然および合成の区別にかかわらず利用することができる。発現の効率は、適切な転写または翻訳のエンハンサー配列を含めることで促進可能である（例えばＢｉｔｔｎｅｒら、Ｍｅｔｈｏｄｓ ｉｎ Ｅｎｚｙｍｏｌ． １５３：５１６、１９８７に記載された配列）。
【００４４】
成分（１）、成分（２）、およびこれらを含む融合タンパク質は、生理学的に正常な条件下で可溶化することができる。またこのような融合タンパク質には、１種または複数の無関係なタンパク質（すなわち非ｈｓｐ、非ＨＰＶのタンパク質）の全体または部分を含めることで、少なくとも３つの部分からなる融合タンパク質を作製することができる。「第３」のタンパク質は、融合タンパク質の精製、検出、または可溶化を促進するタンパク質、または他のいくつかの機能を提供するタンパク質とすることができる。例えば発現ベクターｐＵＲ２７８ （Ｒｕｔｈｅｒら、ＥＭＢＯ Ｊ． ２：１７９１、１９８３）を用いてｌａｃＺ融合タンパク質を作製することが可能であり、またｐＧＥＸベクターを用いて外来ポリペプチドをグルタチオンＳ−トランスフェラーゼ（ＧＳＴ）を含む融合タンパク質として発現させることができる。一般に、このような融合タンパク質は可溶性であり、グルタチオン−アガロースビーズに吸着させてから遊離のグルタチオン存在下で溶出することで溶解細胞から容易に精製することができる。ｐＧＥＸベクターは、クローン化された標的遺伝子産物をＧＳＴ部分と切り離すためにトロンビンまたは第Ｘａ因子プロテアーゼ切断部位を含むように設計されている。「第３」のタンパク質は免疫グロブリンのＦｃドメインでもよい。このような融合タンパク質は親和性カラムで容易に精製することできる。本発明の方法で使用される融合タンパク質が複数の成分（１）および／または複数の成分（２）を含むことはいうまでもなく、成分（１）および（２）を直接または間接的に連結させることができる（例えば一つまたは複数のアミノ酸残基が両者の間に存在するように連結する）。
【００４５】
タンパク質（例えばｈｓｐ、ＨＰＶ抗原、またはｈｓｐを含む融合タンパク質）を、対象タンパク質が特異的に結合する抗体を用いて精製することができる。当業者であれば、親和性に基づく精製法を用いてタンパク質を精製することができる。ヒト細胞系で発現させる非変性型融合タンパク質の精製法については例えばジャンクネート（Ｊａｎｋｎｅｃｈｔ）ら、Ｐｒｏｃ． Ｎａｔｌ． Ａｃａｄ． Ｓｃｉ． ＵＳＡ． ８８：８９７２、１９８１を参照されたい。この系で対象遺伝子は、ワクシニア組換えプラスミドに、遺伝子の読み枠が、６個のヒスチジン残基を含むアミノ末端タグに翻訳的に融合するようにサブクローン化される。組換え体ワクシニアウイルスを感染させた細胞の抽出物をＮｉ２＋ニトリロ酢酸−アガロースカラムに添加し、ヒスチジンタグ付きタンパク質を、イミダゾールを含む緩衝液で選択的に溶出する。これと同じ手法を細菌培養物に使用することができる。
【００４６】
融合タンパク質（特に短い抗原性断片を含むもの）を含むタンパク質を、化学合成（例えば「固相ペプチド（Ｓｏｌｉｄ Ｐｈａｓｅ Ｐｅｐｔｉｄｅ Ｓｙｎｔｈｅｓｉｓ）」、第２版、１９８４、Ｔｈｅ Ｐｉｅｒｃｅ Ｃｈｅｍｉｃａｌ Ｃｏ．、Ｒｏｃｋｆｏｒｄ、ＩＬに記載された方法）で作製することができる。
【００４７】
単離したタンパク質を、望ましいならば、さらに行う処理が本発明の方法で効率的に十分な免疫応答の誘導能力を損なわない限りにおいて、さらに精製および／または濃縮する。タンパク質を精製および濃縮するさまざまな方法は当技術分野で周知である（例えばＦｉｓｈｅｒ、「生物学および分子生物学における実験技術（Ｌａｂｏｒａｔｏｒｙ Ｔｅｃｈｎｉｑｕｅｓ Ｉｎ Ｂｉｏｃｈｅｍｉｓｔｒｙ Ａｎｄ Ｍｏｌｅｃｕｌａｒ Ｂｉｏｌｏｇｙ）」、ＷｏｒｋおよびＢｕｒｄｏｎ編、Ｅｌｓｅｖｉｅｒ、１９８０を参照）。このような方法には例えば超遠心法、および／または沈殿法（例えば硫酸アンモニウムを使用する方法）、微量濾過（例えば０．４５ μｍの酢酸セルロースフィルターを使用する方法）、限外濾過（例えば分粒膜および再循環濾過を用いる方法）、ゲル濾過（例えばセファロース ＣＬ−６Ｂ、 ＣＬ−４Ｂ、 ＣＬ−２Ｂ、６Ｂ、４Ｂ、もしくは２Ｂ、セファクリル Ｓ−４００もしくはＳ−３００、スーパーロース ６またはウルトロゲル Ａ２、Ａ４、もしくはＡ６を充填したカラム；いずれもファルマシアから入手可能）、高速タンパク質液体クロマトグラフィー（ＦＰＬＣ）、ならびに高速液体クロマトグラフィー（ＨＰＬＣ）などがある。
【００４８】
交差反応性 ＨＰＶ 配列
当業者であれば、第１の型のＨＰＶ抗原を含む組成物を用いて、第２の型のＨＰＶに感染した対象を治療可能か否かを判断することができる。このような判断の元となるアッセイ法には、予測アッセイ法（例えばコンピューターモデルを使用した方法）、および生物学的アッセイ法（交差反応性を検討する方法）などがある。一つまたは両方のタイプのアッセイ法を使用することができる（予測アッセイ法で得られた結果を生物学的アッセイ法でさらに検討することは一般的に行われる）。以下にそれぞれの例を説明する。
【００４９】
交差反応性（すなわち一つの型のＨＰＶ抗原を含む組成物が効率的に別の型にＨＰＶに感染した対象、または別の型のＨＰＶに関連した疾患または症状を有する対象を有効に治療する能力）を、十分に確立された免疫学的方法で検討することができる。例えばヒトＭＨＣクラスＩ 分子の抗原結合領域およびヒトＣＤ８遺伝子を発現するように作製した二重トランスジェニック（ｂｉ−ｔｒａｎｓｇｅｎｉｃ）マウス（Ｌｕｓｔｇａｒｔｅｎら、Ｈｕｍａｎ Ｉｍｍｕｎｏｌ． ５２：１０９、１９９７；Ｖｉｔｉｅｌｌｏら、Ｊ． Ｅｘｐ． Ｍｏｄ． １７３：１００７、１９９１）を用いて免疫交差反応性決定することができる。
【００５０】
より具体的には、ＨＬＡ−Ａ２／ＣＤ８二重トランスジェニックマウス（Ｌｕｓｔｇａｒｔｅｎら、前掲）を用いて、ＨＰＶ１６ Ｅ７に対する細胞傷害性Ｔリンパ球（ＣＴＬ）の、ＨＰＶ６および１１のＥ７タンパク質に由来するペプチドに対する交差反応性を、標準的な免疫学的手法（例えばＣｏｌｉｇａｎら編、「免疫学における最新プロトコール（Ｃｕｒｒｅｎｔ Ｐｒｏｔｏｃｏｌｓ ｉｎ Ｉｍｍｕｎｏｌｏｇｙ）」、Ｊｏｈｎ Ｗｉｌｅｙ ＆ Ｓｏｎｓ、１９９９を参照）で示すことができる。簡単に説明すると、マウスをＨｓｐＥ７融合タンパク質（ＢＣＧ Ｈｓｐ６５分子およびＨＰＶ１６ Ｅ７分子を元にしたもの）で７〜２１日の間隔をおいて１〜３回にわたり免疫化する。ＨｓｐＥ７をリン酸緩衝食塩水（ＰＢＳ）に懸濁し、皮下に１匹あたり１ μｇ〜１０００ μｇの用量範囲で投与する。最終ＨｓｐＥ７投与から７日後にマウスを殺し、脾臓を除去し、組織を単一細胞の懸濁液になるまでばらばらにする。ＨＰＶ Ｅ７に特異的なＣＴＬを、ＨＰＶ１６ Ｅ７、ＨＰＶ６ Ｅ７、およびＨＰＶ１１ Ｅ７に由来するＨＬＡ−Ａ２結合ペプチドを１ μＭの濃度で培地に加えて再刺激を行う。この細胞を例えば、各ウェルに異なるペプチドを入れた６ウェルプレート中で再刺激する。コンピューターアルゴリズムでＨＬＡ−Ａ２結合親和性が最高であると予測されたペプチド（例えば１０個のペプチド）を、ＨＰＶ１６、ＨＰＶ６、およびＨＰＶ１１（または他の任意のＨＰＶ型；Ｐａｒｋｅｒら、Ｊ． Ｉｍｍｕｎｏｌ． １５２：１６３、１９９４を参照；このようなアルゴリズムは国立衛生研究所（ＮＩＨ）のＢＩＭＡＳ （Ｂｉｏｉｎｆｏｒｍａｔｉｃｓ ＆ Ｍｏｌｅｃｕｌａｒ Ａｎａｌｙｓｉｓ Ｓｅｃｔｉｏｎ）のウェブサイト（２００１年６月２６日現在におけるウェブサイトはｈｔｔｐ：／／ｂｉｍａｓ．ｄｃｒｔ．ｎｉｈ．ｇｏｖ／）から入手できる）のそれぞれに用いることができる。また別の場合では、他の２種のＨＰＶ遺伝子型に由来する対応ペプチド（ＨＰＶ１６ Ｅ７のペプチド１１〜２０ならびにＨＰＶ６および１１のペプチド１１〜２０）も使用することができる。
【００５１】
インビトロで再刺激してから一定期間（例えば１週間）の後に、ＣＴＬ活性をＨＰＶ１６ Ｅ７、ＨＰＶ６ Ｅ７、およびＨＰＶ１１ Ｅ７に由来するＨＬＡ−Ａ２結合ペプチドで処理したＴ２標的細胞を溶解することで測定することができる。またＨＰＶ１６ Ｅ７、ＨＰＶ６ Ｅ７、およびＨＰＶ１１ Ｅ７に由来するＨＬＡ−Ａ２結合ペプチドを認識する抗原特異的なＴリンパ球は、文献の方法によるＩＦＮ−γ分泌細胞のＥＬＩＳＰＯＴ分析で測定することができる（Ａｓａｌら、Ｃｌｉｎ． Ｄｉａｇｎ． Ｌａｂ． Ｉｍｍｕｎｏｌ． ７：１４５、２０００）。この分析法は、他のＨＬＡ対立遺伝子のトランスジェニックマウスを対象に実施することができる。
【００５２】
または、ＨｓｐＥ７の免疫化で誘導されたＣＴＬのもつ、ＨｓｐＥ７を用いた性器疣贅の治療を受けているヒトＨＬＡ−Ａ２陽性対象における、ＨＰＶ６または１１のＥ７タンパク質に由来するペプチドと交差反応する能力を測定することができる。治療に先立ち、またＨｓｐＥ７による各治療の数日（例えば７日）後に、末梢血単核球細胞（ＰＢＭＣ）を対象（例えばヒト患者）から単離することができる。このような細胞は、蛍光発生性（ｆｌｕｏｒｏｇｅｎｉｃ）ＭＨＣ−ペプチド複合体（四量体、Ａｌｔｍａｎら、Ｓｃｉｅｎｃｅ ２７４：９４、１９９６）を用いて、またはＥＬＩＳＰＯＴ分析（Ａｓａｌら、Ｃｌｉｎ． Ｄｉａｇｎ． Ｌａｂ． Ｉｍｍｕｎｏｌ． ７：１４５、２０００）で分析することができる。細胞は末梢血から直接アッセイし、ユード（Ｙｏｕｄｅ）ら（Ｃａｎｃｅｒ Ｒｅｓ． ６０：３６５、２０００）に記載された手順でインビトロ再刺激によりアッセイすることができる。インビトロにおける再刺激では、２×１０６／ｍｌのＰＢＭＣを、ＲＰＭＩ１６４０中で１０％ ヒトＡＢ血清（ＲＡＢ）およびペプチド（１０ μｇ／ｍｌ）とともに培養する。再刺激用ペプチドはＨＰＶ１６ Ｅ７から得られ、コンピューターアルゴリズム（Ｐａｒｋｅｒら、前掲）で定義されるＨＬＡ−Ａ２結合親和性が最高であると予測されたペプチド（例えば１０個のペプチド）を含むものとすることができる。４日目に２５ユニット／ｍｌのＩＬ−２を含む１ ｍｌのＲＡＢを各ウェルに添加する。６日目に１ ｍｌの培地を１０ ユニット／ｍｌのＩＬ−２を含む１ ｍｌの培地と交換する。７日目に、放射線を照射した自家ＰＢＭＣ （新鮮なものか凍結後に解凍したもの）を３×１０６ 細胞／ｍｌの濃度で、１０ μｇ／ｍｌのペプチドおよび３ μｇ／ｍｌのβ２−ミクログロブリンを含むＲＡＢに再懸濁する。抗原提示細胞を２時間かけて接着させた後に、洗浄して非接着細胞を除き、１〜２×１０６ エフェクター細胞／ｍｌを添加する。９日目に２５ユニット／ｍｌのＩＬ−２を含む１ ｍｌのＲＡＢを各ウェルに添加する。１３日目にウェルの内容物をマルチプレートに分け、培地（１０ ユニット／ｍｌのＩＬ−２を含む）を当初の容量になるまで添加する。１４日目の細胞を使用する。ＦＡＣＳ分析では四量体を文献の方法（Ａｌｔｍａｎら、Ｓｃｉｅｎｃｅ ２７４：９４、１９９６）で調製する。四量体の添加に用いるペプチドは、ＨＰＶ１６、ＨＰＶ６、およびＨＰＶ１１のＥ７分子に由来するＨＬＡ−Ａ２結合ペプチドとする。コンピューターアルゴリズム（Ｐａｒｋｅｒら、前掲）でＨＬＡ−Ａ２結合親和性が最高であると予測されたペプチド（例えば１０個のペプチド）をＨＰＶ１６、ＨＰＶ６、およびＨＰＶ１１のそれぞれに使用する。また別の場合では、他の２種のＨＰＶ遺伝子型に由来する対応するペプチド（ＨＰＶ１６ Ｅ７のペプチド１１〜２０ならびにＨＰＶ ６および１１のペプチド１１〜２０）も使用する。新鮮なＰＢＭＣまたは再刺激後のＰＢＭＣを、ＰＥ標識ＨＰＶ−Ｅ７ペプチド四量体、およびＦＩＴＣ標識抗ＣＤ８抗体で染色し、文献に記載された手順でフローサイトメトリーで分析する。新鮮なＰＢＭＣおよび再刺激後のＰＢＭＣ中に存在する、ＨＰＶ１６ Ｅ７、ＨＰＶ６ Ｅ７、およびＨＰＶ１１ Ｅ７に由来するＨＬＡ−Ａ２結合ペプチドを認識する抗原特異的なＴリンパ球のＥＬＩＳＰＯＴ分析を、文献に記載された方法（Ａｓａｌら、Ｃｌｉｎ． Ｄｉａｇｎ． Ｌａｂ． Ｉｍｍｕｎｏｌ、７：１４５、２０００）で行う。同様に、他のＨＬＡハプロタイプをもつ対象に以上の手法を適用することができる。
【００５３】
また過去にＨｓｐＥ７で処置されたことがなく、ＨＰＶ １６型Ｅ７に由来するＨｓｐＥ７タンパク質またはペプチドでインビトロで刺激されたことのないＨＬＡ−Ａ２陽性の健康被験者に由来するヒトＰＢＭＣが、他の２種のＨＰＶ遺伝子型（６型および１１型）に由来する対応ペプチドで処理した細胞と交差反応する能力を検討することができる。当技術分野で一般的な手順を用いて細胞の刺激および分析を行う。簡単に説明すると、ＰＢＭＣを末梢血から単離し、接着細胞を非接着細胞と分離し、接着細胞を培養して樹状細胞（ＤＣ）を「免疫学における最新プロトコール（Ｃｕｒｒｅｎｔ Ｐｒｏｔｏｃｏｌｓ ｉｎ Ｉｍｍｕｎｏｌｏｇｙ）」（Ｃｏｌｉｇａｎら編、Ｊｏｈｎ Ｗｉｌｅｙ ＆ Ｓｏｎｓ、ｐｐ ７．３２．７〜８、１９９９）に記載された手順で調製する。非接着細胞を９０％ ＦＣＳ／１０％ ＤＭＳＯ中に凍結保存し、後のアッセイ法の各時点で使用する。
【００５４】
刺激する際には、５０ μｇ／ｍｌのＨｓｐＥ７または４０ μｇ／ｍｌの適切なペプチド、および３ μｇ／ｍｌのβ２−ミクログロブリンで２４時間かけて３７℃で５％ ＣＯ２下でＤＣを処理する（Ｋａｗａｓｈｉｍａら、Ｈｕｍａｎ Ｉｍｍｕｎｏｌ． ５９：１、１９９８）。使用するペプチドはＨＰＶ１６、ＨＰＶ６、およびＨＰＶ１１のＥ７分子に由来するＨＬＡ−Ａ２結合ペプチドとする。コンピューターアルゴリズム（Ｐａｒｋｅｒら、前掲）でＨＬＡ−２結合親和性が最高であると予測されたペプチド（例えば１０個のペプチド）をＨＰＶ１６、ＨＰＶ６、およびＨＰＶ１１のそれぞれに使用する。また別の場合、他の２種のＨＰＶ遺伝子型に由来する対応ペプチド（ＨＰＶ１６ Ｅ７のペプチド１１〜２０ならびにＨＰＶ ６および１１のペプチド１１〜２０）も使用する。ＣＤ８＋細胞を、凍結保存された自家非接着細胞から免疫磁気ビーズ（Ｍｉｌｔｅｎｙｉ Ｂｉｏｔｅｃ）を用いた陽性選択で単離する。ペプチド／タンパク質を添加したＤＣに４２００ ラドの放射線を照射し、自家ＣＤ８＋細胞と１：２０の比になるように、例えば０．５ ｍｌのＲＡＢ中に０．２５×１０５ ＤＣおよび５×１０５ ＣＤ８＋細胞、ならびに１０ ｎｇ／ｍｌのＩＬ−７を含む４８ウェルプレートで混合する。７日目と１４日目に同細胞を、自家ペプチドで処理した接着ＡＰＣで再刺激する（Ｋａｗａｓｈｉｍａ ら、Ｈｕｍａｎ Ｉｍｍｕｎｏｌ． ５９：１、１９９８）。培養物に２〜３日毎に１０ Ｕ／ｍｌのｈＩＬ−２を添加する。ＨＰＶ Ｅ７ペプチドに特異的なＴリンパ球を、インビトロ刺激から７日後および１４日後に、蛍光発生性ＭＨＣ−ペプチド複合体（四量体、Ａｌｔｍａｎら、Ｓｃｉｅｎｃｅ ２７４：９４、１９９６）、またはＥＬＩＳＰＯＴ分析（Ａｓａｌら、Ｃｌｉｎ． Ｄｉａｇｎ． Ｌａｂ． Ｉｍｍｉｍｏｌ． ７：１４５、２０００）で分析する。ＦＡＣＳ分析では、四量体を文献の方法（Ａｌｔｍａｎら、Ｓｃｉｅｎｃｅ ２７４：９４、１９９６）で調製する。四量体の添加に使用するペプチドは上述のように、ＨＰＶ１６、ＨＰＶ６、およびＨＰＶ１１のＥ７分子に由来するＨＬＡ−Ａ２結合ペプチドとすることができる。ペプチドに特異的なＴリンパ球を、ＰＥ標識ＨＰＶ−Ｅ７ペプチド四量体、およびＦＩＴＣ標識抗ＣＤ８抗体で染色し、フローサイトメトリーで分析する（Ｙｏｕｄｅら、Ｃａｎｃｅｒ Ｒｅｓ． ６０：３６５、２０００）。ＨＰＶ１６ Ｅ７、ＨＰＶ６ Ｅ７、およびＨＰＶ１１ Ｅ７に由来するＨＬＡ−Ａ２結合ペプチドを認識する抗原特異的なＴリンパ球のＥＬＩＳＰＯＴ分析を文献の方法（Ａｓａｌら、Ｃｌｉｎ． Ｄｉａｇｎ． Ｌａｂ． Ｉｍｍｉｍｏｌ． ７：１４５、２０００）で実施する。同様に、他のＨＬＡハプロタイプをもつ対象に上記手法を適用することができる。
【００５５】
組成物の投与
本発明は、少なくとも１種のＨＰＶタンパク質抗原（例えばＨＰＶタンパク質抗原（もしくはその抗原性断片）、ｈｓｐ（もしくはその免疫刺激性断片）と混合または抱合体を作製したＨＰＶタンパク質抗原、またはＨＰＶタンパク質抗原（もしくはその抗原性断片）およびｈｓｐ（もしくはその免疫刺激性断片）を含む融合タンパク質）を含む組成物を含む。選択的に、これらのタンパク質を、希釈物（例えばＰＢＳ）または重炭酸溶液（例えば０．２４ Ｍ ＮａＨＣＯ３）などの薬学的に許容される担体中に懸濁することができる。有用な担体は投与様式および投与経路を元に、また標準的な薬学的手法で選択することができる。適切な薬学的な担体および希釈物、ならびに使用上薬学的に必要な物質は「レミントン薬科学（Ｒｅｍｉｎｇｔｏｎ’ｓ Ｐｈａｒｍａｃｅｕｔｉｃａｌ Ｓｃｉｅｎｃｅｓ）」に記載されている。アジュバント、例えばコレラトキシン、大腸菌の易熱性エンテロトキシン（ＬＴ）、リポソーム、または免疫刺激複合体（ＩＳＣＯＭ）も含めることができる。
【００５６】
タンパク質（例えば融合タンパク質）を対象に直接投与する必要はなく、代わりにタンパク質をコードする核酸配列を投与することができる。このようなタンパク質は対象でインビボで発現する。核酸は、（ウイルスベクター、例えばウイルスベクターゲノムなどの）ベクターの一部とすることができるほか、例えばリポソームに包むことができる。核酸を裸の核酸として投与することもできる。
【００５７】
組成物は、溶液、懸濁液、坐剤、錠剤、顆粒、粉末、カプセル、軟膏、またはクリームとして製剤化することができる。上述したように、上記組成物を調製する際には１種または複数の薬学的担体を含めることができる。薬学的に許容される担体もしくは他の付加物の例には溶媒（例えば水もしくは生理食塩水）、可溶化剤（例えばエタノール、ポリソルベート、もしくはクレモフォア ＥＬ （登録商標））、等張性を調節する薬剤、保存剤、抗酸化剤、賦形剤（例えば乳糖、デンプン、結晶セルロース、マンニトール、マルトース、リン酸水酸カルシウム、軽質無水ケイ酸、もしくは炭酸カルシウム）、結合剤（例えばデンプン、ポリビニルピロリドン、ヒドロキシプロピルセルロース、エチルセルロース、カルボキシメチルセルロース、もしくはアラビアゴム）、潤滑剤（例えばステアリン酸マグネシウム、タルク、もしくは硬化油）、または安定剤（例えば乳糖、マンニトール、マルトース、ポリソルベート、マクロゲル、もしくはポリオキシエチレン硬化ヒマシ油）などがある。必要であれば（または望ましくは）、グリセリン、ジメチルアセタミド、乳酸ナトリウム、界面活性剤、水酸化ナトリウム、エチレンジアミン、エタノールアミン、炭酸ナトリウム、アルギニン、メグルミン、またはトリスアミノメタンを使用することができる。ポリ−Ｄ，Ｌ−ラクチド−コ−グリコライドまたはポリグリコライドなどの生分解性ポリマーを、組成物の徐放が望ましい場合には原体マトリックス（ｂｕｌｋ ｍａｔｒｉｘ）として使用することができる（例えば米国特許第５，４１７，９８６号、第４，６７５，３８１号、および第４，４５０，１５０号を参照）。溶液、錠剤、顆粒、またはカプセルなどの薬学的調製物を上記組成物を用いて作ることができる。組成物を経口投与する場合は香料および着色料を添加することができる。
【００５８】
治療的組成物は適切な経路で、例えば経静脈的、経動脈的、局所的、腹腔内、胸膜内、経口的、皮下、筋肉内、皮内、舌下、表皮内、経鼻的、肺内（例えば吸入）、経膣的、または経直腸的に投与することができる。
【００５９】
投与する組成物の量は、例えば特定の組成物、アジュバントが組成物と同時投与されるか否か、同時投与されるアジュバントの種類、投与の様式および回数、ならびに所望の効果（例えば予防または治療）によって変動する。用量は当業者によって、薬剤または予防薬を開発する過程で日常的な手順で決定される。一般に本発明の組成物は、ヒト成人に対する用量として１ μｇ〜１００ ｍｇの量が投与される。アジュバントを組成物とともに投与する場合は、ヒト成人１人に１ ｎｇ〜１ ｍｇの量を一般的に使用することができる。投与は、当業者によって決定されるように必要に応じて繰返し行う。例えば初回投与の後に週に１回、または月に１回の間隔で３回のブースター投与を行うことができる。ブースター投与は初回投与後の３〜１２週の時点で行うことができる。また２回目のブースター投与は、同じ剤形を用いて３〜１２週後に行うことができる。血清またはＴ細胞を対象から採取し、例えば融合タンパク質、またはタンパク質抱合体に含まれるＨＰＶ抗原に対する組成物で生じる免疫応答の検討に用いることができる。特定の抗原に対する抗体または細胞傷害性Ｔ細胞を調べる方法は当技術分野で周知である。付加的なブースター投与を必要に応じて実施することができる。例えば組成物に含まれる融合タンパク質の量を変化させることで、最大の免疫応答を誘導するように免疫化のプロトコルを最適化することができる。
【００６０】
本明細書に記載されたタンパク質を、ウイルスベクター（例えばレトロウイルスまたはアデノウイルスのベクター）などの核酸を用いて投与できることはいうまでもない。
【００６１】
さらに詳細に記述することはしないが、当業者であれば上記の記述および以下の実施例をふまえて本発明を十分使用できるものと思われる。以下に挙げる実施例は、当業者が融合ポリペプチドを単離して使用することを単純に説明するためであると解釈され、開示の残りの部分をどんな形にせよ制限しない。
【００６２】
実施例
ＨＰＶ １６型のＥ７タンパク質を結合させたウシ型結核菌ＢＣＧ Ｈｓｐ６５を含む融合ポリペプチドを組換え的に作製し、国際公開公報第９９／０７８６０号に記載された手順で製剤化した。Ｈｓｐ６５はストレスタンパク質のＨｓｐ６０ファミリーの分子である。肛門の重度の扁平上皮内病変（ＨＳＩＬ）の治療におけるこの融合ポリペプチドの有効性を検討する、ヒトを対象とした臨床試験の過程で以下の観察がなされた。
【００６３】
肛門ＨＳＩＬ治療におけるＨｓｐＥ７の無作為化した、二重盲の偽薬対照多施設試験には、２２人の患者が参加した。適格患者は生検で肛門ＨＳＩＬであることが確認され、ヒト免疫不全ウイルス（ＨＩＶ）の検査結果は陰性であった。患者のＨＰＶ型を肛門検査で得られた細胞を用いて決定したが、ＨＰＶ−１６を有する必要はなかった。個々の病変のＨＰＶ型は決定しなかった。患者は１００ μｇのＨｐｓＥ７または偽薬の３回の皮下注射を毎月受けた。また肛門の乳頭腫コロニー試験、生検を伴う高分解能の肛門鏡検査（ＨＲＡ）、および医師による全身評価で治療反応の評価が行われた。対照試験で１２週または２４週後に非反応者（肛門ＨＳＩＬが持続している者）はオープンラベル試験に変更させた。オープンラベル試験では、患者は５００ μｇのＨｓｐＥ７の ３回の注射を毎月受けた。治療の評価は偽薬対照試験における二重盲検とし、処置内容は開示されていない。
【００６４】
患者患者のＨＰＶ型を決定するためにダクロン（Ｄａｃｒｏｎ）スワブを用いて、無作為化偽薬対照試験のスクリーニング受診時に患者の肛門から検体を生検直前に採取した。試料運搬培地（Ｓａｍｐｌｅ Ｔｒａｎｓｐｏｒｔ Ｍｅｄｉｕｍ）（Ｄｉｇｅｎｅ）に移した後にＤＮＡを単離してＨＰＶ型の決定に用いた。簡単に説明すると、コンセンサスプライマーセットＭＹ０９／ＭＹ１１を用いてＨＰＶ ＤＮＡをポリメラーゼ連鎖反応（ＰＣＲ）で増幅した。増幅段階に続いて試料をナイロン膜上にブロットし、２９種のＨＰＶ型
【００６５】
ＰＣＲが成功したことを検証するために、βグロビン対照の増幅およびプローブによる検出を各試料を対象に行った。試料がβグロビンの存在に関して陽性を示さなかった場合、ＰＣＲ段階は技術的に失敗したとみなした。コンセンサスプローブによって２段階またはそれ以上の評点が得られなかった場合、対象試料は「ＨＰＶ陰性」とみなした。
【００６６】
オープンラベル試験へのエントリーの時点で、２２人中１４人の患者（６４％）で、１００ μｇのＨｓｐＥ７または偽薬を毎月３回注射された、事前の二重盲試験を通して、持続した肛門性器疣贅が認められた。この１４人の患者のうち、オープンラベル試験への切り換え時点までに、８人（５７％）に悪化が認められ、４人の患者（２９％）に変化が認められず、また２人の患者（１４％）に改善が認められた（１人は劇的に改善し、もう１人はわずかに改善した）。別の１人の患者には二重盲試験の開始時点でコンジロームが認められたが、オープンラベル試験開始前に解消し、いずれの試験中にも疣贅が検出されなかった他の７人とともに分析対象から除外された。コンジロームは１４人の患者全員（１００％）の肛門直腸管内部に認められ、１４人中６人（４３％）では肛門周囲の皮膚にも認められた。治験担当医は、オープンラベル試験の開始時点で疣贅の存在が認められた１４人の患者のうち、外科的切除が１１人の患者（７９％）に、局所切除（例えば液体窒素または電気焼灼）が２人の患者（１４％）に、また局所療法（イミキモド（ｉｍｉｑｕｉｍｏｄ））が１人の患者（７％）にそれぞれ必要であると判断した。これらの患者は、治験担当医が推奨する治療の延期を選択し、代わりにオープンラベル試験でＨｓｐＥ７ ５００ μｇの３回の注射を毎月受けることに同意した。
【００６７】
５００ μｇのＨｓｐＥ７を用いた最後の治療から１か月後、２人の患者（１４％）に検出可能な疣贅は認められず、１１人の患者（７９％）にオープンラベル試験エントリー時の状況と比較して疣贅の大きさの低下または数の減少が認められ、また１人の患者（７％）に疣贅の大きさの拡大が認められた（表１）。オープンラベル試験の初期評価時点（最終投与から４か月後）までに、さらに１人の患者に有効から著効の改善が認められた（視認可能な疣贅が確認されない）。このため合計３人（２１％）が著効を示したことになる（表１）。これらの反応者ではいずれもオープンラベル試験の６か月の評価期間中に再発は認められなかった。１０人の患者（７１％）は有効な改善を継続して示した（すなわち、疣贅の大きさがさらに有意に縮小し、残存する疣贅の除去に必要な治療の程度が継続して低下した）。１人の非反応者（７％）では、オープンラベル試験の終了時までに改善が認められなかった
【００６８】
【表１】ＨｓｐＥ７を用いた治療後の肛門性器疣贅の反応一覧
†５００ μｇのＨｓｐＥ７を用いた最終治療から４か月後
【００６９】
試験終了時に治験担当医は、著効を示した３人の対象に対して治療の続行を推奨しなかった。表２に示したように、結果が有効であった者に対する治験担当医の推奨する治療法の内訳は、除去療法（１４人中６人、４３％）、または局所薬剤を用いた治療（１４人中４人、２９％）であった。また非反応者には追加手術が勧められた（１４人中１人、７％）。全２２人が反応の長期追跡調査に対してレジストリプロトコル（ｒｅｇｉｓｔｒｙ ｐｒｏｔｏｃｏｌ）に進み、担当医の推奨する治療を延期することに同意した。
【００７０】
【表２】肛門性器の反応の評価および医師が推奨する治療法
†５００ μｇのＨｓｐＥ７を用いた最終治療から１か月後
‡５００ μｇのＨｓｐＥ７を用いた最終治療から４か月後
略号：ＣＲ＝著効；ＰＲ＝有効
【００７１】
肛門性器疣贅と診断された全１４人中、複数のＨＰＶ型のＨＰＶ ＤＮＡが、最初の無作為化対照試験のスクリーニング中に肛門塗抹標本で検出された（表３）。ＨＰＶ−６および／または１１は、１２人の患者（８６％）で認められた。１人の患者ではＨＰＶ−１６および関連型のみが認められ、他の患者では型は確定できなかった。１４人の患者中３人（２１％）がＨＰＶ−１６陽性であった。疣贅の改善がみられた大部分の患者（１３人中１１人、８５％）にはＨＰＶ−１６は認められなかった。非反応者でもＨＰＶ−１６は認められなかった（表３参照）。
【００７２】
【表３】肛門性器疣贅患者のＨＰＶ型
†５００ μｇのＨｓｐＥ７を用いた最終治療から４か月後
略号：ＨＰＶ＝ヒト乳頭腫ウイルス；ＣＲ＝著効；ＰＲ＝有効
【００７３】
ＨｓｐＥ７ （５００ μｇを３か月毎）を投与した、持続性の肛門ＨＳＤＬ患者および随伴性肛門性器疣贅がみられる患者を対象としたオープンラベル交差試験では、基線時に疣贅が認められた１４人中３人の患者（１７％）では、最終治療から４か月後に疣贅は認められなかった。別の１０人の患者（７１％）には症状に改善が認められた（疣贅の大きさが有意に縮小し、残存する疣贅の除去に必要な治療の程度が継続して低下した）。１人の患者（７％）では試験中に改善が認められなかったので、治験担当医により追加手術が推奨された。
【００７４】
オープンラベル試験の登録前に、治験実施施設の大部分の患者は、疣贅除去に対する外科的介入を受けていた（１４人中１１人、７９％）。試験終了時までに外科的治療が推奨されたのは１人のみであった。局所除去療法（例えば液体窒素、電気焼灼）が６人の患者（４３％）に推奨され、局所薬剤（例えばイミキモド）を用いた治療が４人の患者（２９％）に推奨された。３人の患者は後の治療を必要としなかった。
【００７５】
６か月間における反応は進行性とみなされ、同期間に再発した反応者はいなかった。コンジロームの漸進的および進行的な解決は、ＨｓｐＥ７による細胞性免疫の誘導後における免疫学的宿主反応に基づく予測と違わない。
【００７６】
二重盲試験に参加した２人の患者では、オープンラベル試験に進む前にコンジロームにある程度の改善が認められた。現時点において発明者らは無作為化の内容を知らされておらず、これらの患者が１００ μｇのＨｓｐＥ７または偽薬のいずれの投与を受けたかのは不明である。しかし５００ μｇのＨｓｐＥ７を用いた毎月３回の注射を行ったオープンラベル試験で観察された反応に基づき、高用量の方が１００ μｇの場合と比べて活性が高いと考えられる。
【００７７】
ＨＰＶ−１６のＤＮＡは、ＨｓｐＥ７による治療後に疣贅の改善がみられた１３人中３人の患者（２３％）のみの肛門塗抹標本で検出された。ＨＰＶ−６、ＨＰＶ−１１、または両方に由来するＤＮＡは、疣贅がＨｓｐＥ７を用いた治療に反応した患者の大部分で検出された。以上のデータは、これらのＨｓｐＥ７に対する反応において上記ＨＰＶ型の間に免疫学的な交差反応が存在することを示唆している。
【００７８】
要約すると、本明細書に示した結果は、ＨｓｐＥ７が肛門性器疣贅に対して広範囲に活性があることを示唆している。この活性はＨＰＶ−１６陽性患者に制限されないようであるが複数のＨＰＶ型と交差する。ＨｓｐＥ７は、ＨＰＶに起因する陰部疾患の治療に活性を示し、この活性はＨＰＶ−１６陽性患者に制限されないと考えられる。
【００７９】
本明細書に報告された観察は、ＨｓｐＥ７を用いた治療が、肛門性器疣贅に対する新しくて単純かつ非外科的な治療法となる可能性があることを示唆している。そして少なくとも疣贅による負荷を緩和することで治療の程度、および結果として生じる罹患率を低下させる可能性がある。肛門直腸内部の疾患は、かなりの痛みを伴う消耗性の追加的治療を必要とする場合がある。「外科的」療法すなわち除去療法の量または程度を低下させたり、または無くす、部分的な反応をもたらす程度の治療法も、罹患率の低下、作業時間の短縮、およびＱＯＬの改善につながる。
【００８０】
以上の結果は、熱ショックタンパク質／ＨＰＶ １６型抗原組成物が、主にＨＰＶ ６型および１１型に起因すると考えられる疣贅の、除去または縮小に有効なことを示している。重要な点は、（１）疣贅がＨＰＶに基づく組成物のすべてで治療可能であり、また（２）ＨＰＶ １６型の組成物が、１６型以外のＨＰＶに起因すると考えられる状態の治療に有効である、という結果である。後者の交差反応の結果は、型特異的な組成物は型特異的な免疫応答のみを生じるという一般的な理解をふまえれば、ほとんど予測できない結果であった。
【００８１】
融合ポリペプチドについて観察された交差反応性の機構を詳しく調べるために、理論上の結合を、さまざまなＨＬＡクラスＩ分子およびＨＰＶ １６型、６型、および１１型のＥ７ペプチドについて算出した。解離のＴ１／２はパーカー（Ｐａｒｋｅｒ）ら、Ｊ． Ｉｍｍｕｎｏｌ． １５２：１６３、１９９４ （上述のウェブサイトも参照）に記載されたアルゴリズムを用いて算出した。データを表４にまとめる。
【００８２】
【表４】
【００８３】
表４の結果から、調査対象の特定のＨＬＡ分子に依存して、ＨＰＶ １６型のＥ７抗原が他のＨＰＶ型のＥ７抗原に対する細胞性免疫応答を引き起しうることが示唆される。例えばＨＬＡ Ｂ ２７０５の場合、全３種のＨＰＶ型のＥ７のアミノ酸７６位から始まるペプチドについて、高レベルの結合の存在が予測された。したがって同ＨＬＡ分子を発現する患者では、ＨＰＶ １６型のＥ７組成物は交差反応し、またＨＰＶ ６型および１１型の感染の治療または予防に有用であると考えられる。表４の太字で記載された各ペプチド断片は、完全なＥ７ウイルス抗原に対する基質として組成物（例えば本明細書に記載された融合ポリペプチド）に含めることが可能な抗原性断片である。２種またはそれ以上の推定ＨＬＡエピトープ、または多くの推定ＨＬＡエピトープを含む長い断片も使用可能であることはいうまでもない。[0001]
Field of the invention
The present invention relates to a method for treating human papillomavirus infection.
[0002]
Background of the Invention
Human papillomavirus (HPV) infection is prevalent. HPV is transmitted sexually and is estimated to affect 20-80% of sexually active adults. Most infections are asymptomatic, but infections can lead to the development of genital warts (the incidence is about 1-5% of adults) and the development of genital cancer. Cervical cancer, another type of cancer, is strongly associated with HPV (Frazer, Genitourin. Med.72: 398-403, 1996). HPV types 6, 11, 16, 18, 31, and 33 may be associated with an increased risk of cancer, with type 16 and / or 18 being more than 90% of cervical cancers. In many (van Driel et al., Ann. Med.28: 471-477, 1996). Types 6 and 11 are also associated with anogenital warts. For a review on papillomaviruses and related lesions, see Shah et al., "Chapter 66: Papillomaviruses," Virology, 3rd Edition, edited by Fields et al., Raven Press, Philadelphia, pp 2077-2109, 1996), and zur Hausen, J. Mol. Natl. Cancer Inst.92: 690-698, 2000.
[0003]
Currently, there is no safe and effective way to treat or prevent warts or the above diseases by targeting the immune system. Efforts to develop such treatments have been stalled for several reasons. One reason is the established belief that antigens from one HPV type produce only a limited type-specific immune response. Thus, it has been suggested that cocktails containing antigens from a wide variety of HPV types are required for broadly effective HPV therapy (Caine et al., Science).288: 1753, 2000).
[0004]
Summary of the Invention
The present invention is based, in part, on the discovery that a fusion protein comprising a protein from one HPV type can be used to treat a disease or condition caused by infection of another HPV type. For example, HPV type 16 antigen fused to bacterial heat shock protein (hsp) is effective in treating human genital warts caused by HPV types other than type 16 (eg, HPV types 6 and 11). there were. The results support two points: (1) that warts can be treated with HPV proteins, and (2) therapeutics for HPV are derived from different HPV types for the purpose of broad spectrum efficacy. It does not need to contain protein antigens.
[0005]
Accordingly, the present invention provides a subject comprising (1) hsp, or an immunostimulatory fragment thereof, and (2) an HPV protein (eg, an antigenic protein such as the HPV 16 type E7 protein), or a composition comprising the antigenic fragment thereof. The method of treating a wart in a subject. Such compositions will be referred to herein as "Component (1)" and "Component (2)," respectively. The hsp (or immunostimulatory fragment thereof) and the HPV protein (or antigenic fragment thereof) can be simply mixed in the same preparation or tightly bound by chemical conjugation or fusion (ie, the present invention). A fusion protein comprising the components described herein, or a nucleic acid encoding it, can be administered). The components (1) and (2) can be administered simultaneously by mixing, conjugation or fusion. However, each component can be administered separately (eg, continuously) or component (2) can be administered without component (1). The methods described above can include the step of identifying a subject having a wart or suspected of having a wart (in the sense of treating a subject, the identification is made prior to the start of administration of the therapeutic agent). In some cases). Physicians and other persons skilled in the art can appropriately identify such patients.
[0006]
The methods of the present invention can also be used to prevent warts when a subject who wishes to prevent warts (or is not already having warts) or who will benefit from wart prevention is identified.
[0007]
The present invention also provides administering to a subject a composition comprising (1) hsp, or an immunostimulatory fragment thereof, and (2) a second type (eg, type 16) protein of HPV, or an antigenic fragment thereof. A disease caused by an HPV infection of a first type (eg, type 5, 6, type 11, type 18, type 31, type 33, type 35, type 45, type 54, type 60 or type 70) or It features a method of treating a subject having a condition. The “first type” HPV and the “second type” HPV are different from each other and are two different types of HPV. hsp (or an immunostimulatory fragment thereof), and HPV protein (or an antigenic fragment thereof) can be simply mixed together in the same preparation or more tightly bound by chemical conjugation or fusion (Ie, a fusion protein having the components described herein, or a nucleic acid molecule encoding the same, can be administered). The components (1) and (2) can be administered simultaneously by mixing, conjugation or fusion. On the other hand, each component can be administered separately (for example, continuously), or component (2) can be administered without component (1). Further, such methods can include identifying a subject infected with, or suspected of having, an infection (or associated disease or condition).
[0008]
By administering to a person infected with a first type of HPV a composition comprising a second type of HPV, the method described above can be used to classify the HPV infection before it becomes manifest, It can be performed before appearing (ie, it is not necessary to know the particular HPV type that the subject is infected or at risk of becoming infected before treatment or prevention begins). Where the method is prophylactic, the method can include the step of identifying a subject who desires or will benefit from prevention of HPV infection.
[0009]
The compositions described herein may be used (e.g., by reducing the size of a wart or changing the shape of a wart, or by wart-related symptoms (e.g., pain often associated with plantar warts). If the subject has more than one wart, the treatment can include reducing the number of warts if the subject has multiple warts. . The compositions described herein may also be used for diseases caused by or associated with HPV infection (eg, cancers such as cervical or anal cancer, or cervical or anal dysplasia, for example). And other conditions). Although warts are referred to individually as described above, warts can include conditions that are attributable to HPV or conditions that are associated with HPV. Physicians and other persons skilled in the art will understand a "treatment" that is effective for a disease or condition associated with a wart or HPV when the undesirable physiological effects associated with the wart or disease or condition are reduced. . For clinical and physiological symptoms of warts, and clinical and physiological symptoms of diseases or conditions associated with HPV infection, see, eg, Fauchi et al., “Harrison's Principles of Internal Medicine (Harrison's Principles of Internal). Medicine), 14th edition, McGraw-Hill Press, New York, pp. 302-303 and 1098-1100, 1998.
[0010]
A “subject” that would benefit from the methods described herein is an animal (eg, a mammal, such as a human or livestock (eg, cows, horses, pigs, sheep, and goats) that can be infected with papillomavirus, and a livestock animal. An animal (eg, a cat or dog) is a wart in the genital, skin, or internal organs of a subject (also called recurrent respiratory papillomatosis (RRP; juvenile laryngeal papillomatosis (JLP) or adult-onset RRP)) Warts that appear in the vocal cords).
[0011]
The present invention is for use in treating a subject having a wart or disease or condition associated with (or due to) HPV infection (protein, protein conjugate) according to the methods described herein. And / or fusion proteins, or nucleic acid molecules encoding them) further comprising the use of one or more of the compositions described herein. The invention also relates to the use of one or more such drugs in the manufacture of a medicament for treating a subject having a disease or condition associated with (or due to) a wart or HPV infection according to the methods described herein. It also includes the use of the composition.
[0012]
An “antigenic fragment” of a protein (eg, an HPV protein) is intended to target a fragment-specific or protein-specific immune response from which the fragment is derived to the protein when administered in the manner described herein. Any part of the protein that occurs in The immune response can be either a humoral or a cellular response. For example, the antigenic fragment may be an HLA class I peptide antigen described below. Those skilled in the art will appreciate that the immune response desired in the context of the present invention not only occurs with intact proteins and protein fragments, but also with mutated proteins (e.g., the addition, substitution (e.g. It will be understood that this can also occur with amino acid substitutions) or proteins with deletions). It is easy to generate mutant HPV antigens and to study their effects in the context of the present invention.
[0013]
An “immunostimulatory fragment” of a protein (eg, hsp) is any part of a protein that, when administered in accordance with the present invention, promotes an immune response by an antigen. For example, if the immune response to the HPV protein is enhanced when the HPV protein is administered (eg, in a fused state) with a fragment of hsp, such fragment is an immunostimulatory fragment of hsp. One of skill in the art will appreciate that such an immune response may be enhanced by a mutant hsp (eg, an hsp containing an addition, substitution (eg, a conservative amino acid substitution) or deletion at one or more positions in the amino acid sequence). I think you will understand. It is easy to prepare the mutant hsp and examine its ability to promote an immune response to HPV antigens.
[0014]
The method of the present invention provides an efficient means of: (1) treating or preventing warts, and (2) with compositions comprising (ie, using compositions of) other types of HPV. ), Treating or preventing a disease or condition caused by (or associated with) an infection of an HPV type. Thus, compositions comprising HPV antigens of one HPV type can be used in many if not most subjects, regardless of the infected (or potentially infected) HPV type. It is surprising that HPV compositions exhibit efficacy in such environments (environments requiring cross-reactivity). It has previously been considered that one type of HPV antigen does not produce an effective immune response against another type. Other features or advantages of the invention will be apparent from the following detailed description, and from the claims.
[0015]
Detailed description
The present invention relates to broadly effective HPV-based therapeutics, including hsp and HPV proteins (eg, protein antigens). Without limiting the present invention to HPV-based therapies that act via a particular mechanism, such agents can be used to control warts and other conditions associated with HPV infection (eg, dysplasia), and diseases (eg, Cancer). For example, a composition of the present invention may include HPV proteins derived from more than one HPV type, but may include HPV proteins derived from only one type. In addition, compositions comprising HPV proteins from one HPV type are useful for treating or preventing warts or other diseases or conditions associated with HPV resulting from other types of HPV infection. Various materials and procedures suitable for use in connection with the present invention are described below.
[0016]
Preparation of fusion protein
Nucleic acid sequences encoding hsp and HPV proteins are known and utilized by those skilled in the art. Thus, nucleic acid constructs encoding fusion polypeptides useful in the methods of the present invention can be readily prepared in a routine manner (also such nucleic acid molecules can be used separately for the production of hsp and HPV proteins). In addition to being able to be used, the individual hsp and HPV proteins can be physically linked later (eg, simply by mixing), linked by chemical conjugation (described below), or via disulfide bonds. it can). Examples of nucleic acid sequences encoding hsp selectively fused to an antigen (eg, an HPV antigen) are described in WO 89/12455, 94/29459, 98/23735, and 99/07860. , As well as the references cited herein. A method for expressing and purifying a protein containing a fusion protein is described below.
[0017]
Preparation of protein conjugate
Component (1) and component (2) can also be linked by post-translational binding between each hsp and each HPV antigen. Methods for chemically linking two proteins (or fragments thereof) are well known in the art (eg, Hermanson, "Bioconjugate Technologies", Academic Press, San Diego, CA, 1996; Lussow). Eur. J. Immun.21: 2297-2302, 1991; and Barrios et al., Eur. J. Immun.221365-1372, 1992). The conjugate can be prepared by a method using a cross-linking agent such as glutaraldehyde (which becomes a part of the resulting conjugate) or by a method in which component (1) and component (2) are linked by a disulfide bond. . The formation of intermolecular disulfide bonds can also be promoted using cysteine residues naturally present in hsp or cysteine residues inserted recombinantly, or both. Compositions comprising hsp, or an immunostimulatory fragment thereof (component (1)) that binds non-covalently to an HPV antigen are described in US Pat.
[0018]
Regardless of the final form of the dosage composition, component (1) and component (2) may include the molecules listed below.
[0019]
HPV Protein antigen
Any HPV antigen is suitable for use in the compositions of the invention (eg, the mixtures, conjugates, and fusion proteins described herein). However, HPV antigens that express a recognizable epitope on the surface of HPV-infected cells should be particularly useful. HPV expresses six or seven nonstructural proteins and two structural proteins, both of which act as targets in the immunological prophylaxis or immunological therapies described herein. I do.
[0020]
The viral capsid proteins L1 and L2 are late structural proteins. L1 is a major capsid protein whose amino acid sequence is well conserved in various HPV types. There are also seven primary nonstructural proteins. Proteins El, E2, and E4 play important roles in viral replication. Protein E4 also plays a role in virus maturation. The role played by E5 is not well understood. Proteins E6 and E7 are oncoproteins essential for viral replication and host cell immortalization and transformation.
[0021]
Hsp
Various hsps from a variety of species have been isolated, cloned, and characterized (Mizzen, Biotherapy).10173-189, 1998; as used herein, the term "heat shock protein", or its abbreviation hsp, is synonymous and includes proteins referred to as "stress proteins." Hsp, or an immunostimulatory fragment thereof, that stimulates the immune system is suitable for use in the compositions described herein (eg, a portion of a fusion polypeptide). hsp70, hsp60, hsp20-30, and hsp10 are a major group of determinants recognized by the host immune response to infection by Mycobacterium tuberculosis and Mycobacterium leprae. In addition, hsp65 of Bacillus Calmette Guerin (BCG), which is one strain of M. bovis, has been found to be an effective immunostimulant as described in Examples.
[0022]
The hsp gene and hsp family can be used as described for component (1) and are well known in the art. This family includes, for example, Hsp100-200, Hsp100, Hsp90, Lon, Hsp70, Hsp60, TF55, Hsp40, FKBP, cyclophilin, Hsp20-30, ClpP, GrpE, Hsp10, ubiquitin, calnexin, and protein disulfide isomerase. This is described, for example, in Macario, Cold Spring Harbor Laboratory Res.25: 59-70, 1995; Parsell et al., Rev .. Genet.27: 437-496, 1993; and U.S. Patent No. 5,232,833. The hsp may be, but is not limited to, a mammalian, bacterial, or mycobacterial hsp.
[0023]
Grp170 (glucose-regulated protein) is an example of an hsp in the hsp100-200 family. Grp170 is present in the lumen of the endoplasmic reticulum, the pre-Golgi apparatus, and may play an important role in immunoglobulin folding and assembly.
[0024]
Examples of hsp from the hsp100 family include mammalian Hsp110, yeast Hsp104, and E. coli hsp (ClpA, ClpB, ClpC, ClpX, and ClpY).
[0025]
Examples of the hsp90 family include E. coli HtpG, yeast Hsp83 and Hsc83, and human Hsp90α, Hsp90β, and Grp94. Hsp90 is a class of proteins that are normally cell regulatory molecules, such as steroid hormone receptors (eg, glucocorticoid, estrogen, progesterone, and testosterone receptors), transcription factors, and protein kinases that play important roles in signal transduction. Join. The Hsp90 protein is also involved in the formation of large and abundant protein complexes, including other stress proteins.
[0026]
Lon is a tetrameric ATP-dependent protease that degrades E. coli unnatural proteins.
[0027]
Examples of hsp of the hsp70 family include Hsp72 and Hsc73 in mammalian cells, bacteria or mycobacteria (Ray, M. tuberculosis, and M. bovis (such as Calmette-Guerin bacillus; described herein as hsp71); , Yeast, and other prokaryotes, such as DnaK, and BiP and Grp78.hsp70 can specifically bind to ATP, and unfolded polypeptides and peptides.hsp70 can fold and unfold proteins, And involved in the assembly and degradation of protein complexes.
[0028]
An example of an Hsp60 family hsp is the mycobacterial Hsp65. Bacterial Hsp60 is also widely known as GroEL. Hsp60 forms a large homo-oligomer complex and is thought to play an important role in protein folding. Hsp60 homologs are present in eukaryotic mitochondria and chloroplasts.
[0029]
Examples of hsps of the TF55 family include Tcp1, TRiC, and thermosome. These proteins are usually found in the cytoplasm of eukaryotes and some archaebacteria, forming multi-membered rings that facilitate protein folding. They also have weak homology to Hsp60.
[0030]
Examples of hsps from the Hsp40 family include DnaJ, a prokaryotic organism such as E. coli and mycobacteria, and HSJ1, HDJ1, and Hsp40. Hsp40 acts as a molecular chaperone in cellular activities such as protein folding, thermotolerance, and DNA replication.
[0031]
Examples of FKBP include FKBP12, FKBP13, FKBP25, FKBP59, Fpr1, and Nep1. These proteins usually have peptidyl-propyl isomerase activity and interact with immunosuppressants such as FK506 and rapamycin. These proteins are usually present in the cytoplasm and endoplasmic reticulum.
[0032]
Examples of cyclophilins include cyclophilins A, B, and C. These proteins have peptidyl-prolyl isomerase activity and interact with the immunosuppressant cyclosporin A.
[0033]
Hsp20-30 are also called low molecular weight Hsp (small Hsp). Hsp20-30 is usually present in large homo-oligomer complexes and may be found in hetero-oligomer complexes. Depending on the type of organism or cell, a plurality of different types of low molecular weight hsp may be expressed. Hsp20-30 may interact with cytoskeletal structures and play a role in regulating actin polymerization / depolymerization. Hsp20-30 are rapidly phosphorylated when stressed or resting cells are exposed to growth factors. Hsp20-30 homologs include α-crystallin.
[0034]
ClpP is a protease from E. coli that is involved in abnormal protein degradation. Homologs of ClpP are present in chloroplasts. ClpP forms a hetero-oligomer complex with ClpA.
[0035]
GrpE is an approximately 20 kDa Escherichia coli protein involved in the rescue of stress-damaged proteins as well as the degradation of damaged proteins, and GrpE plays an important role in regulating the expression of stress genes in E. coli.
[0036]
Examples of Hsp10 include GroES and Cpn10. Hsp10 is found in E. coli and in mitochondria and chloroplasts of eukaryotic cells. Hsp10 forms a seven-membered ring that binds to the Hsp60 oligomer. Hsp10 is also involved in protein folding.
[0037]
Ubiquitin has been found to bind to proteins that regulate the proteolytic removal of proteins by ATP-dependent cytoplasmic proteases.
[0038]
Stress proteins useful in the present invention include enterobacteria (e.g., E. coli), mycobacteria (especially Lycobacterium tuberculosis, Mycobacterium tuberculosis, Mycobacterium baccae, M. vaccae, and Mycobacterium bovis). Obtained from yeast, Drosophila, vertebrates (eg, birds or mammals such as rodents or primates, including humans).
[0039]
Protein expression and purification
The protein can be made recombinantly. Specifically, a fusion protein comprising hsp (or a fragment thereof) and an HPV antigen (or a fragment thereof), which can be administered individually, in combination, or administered after conjugation, and a fusion protein containing component (1) and component (2), , Yeast, plants or plant cells, or animals or animal cells. For example, hsp, HPV antigens, and fusion proteins containing them can be produced in host cells by transformation (transfection, transduction, or infection) using nucleic acid sequences on a suitable expression vehicle. Suitable expression vehicles include plasmids, viral particles, and phages. Baculovirus expression vectors are suitable for insect cells. All or part of the expression vehicle can be integrated into the host cell genome. In some situations, it may be desirable to use an inducible expression vector, such as the LACSWITCH® "Inducible Expression System" (Stratagene; La Jolla, CA).
[0040]
One of skill in the art of molecular biology will understand that any of a variety of expression systems can be used to provide recombinant proteins (eg, fusion proteins) useful in the methods described herein. The exact host cell and vector used is not critical to the invention.
[0041]
As described above, component (1), component (2), and a fusion protein containing them can be produced in plant cells. For plant cells, viral expression vectors (eg, cauliflower mosaic virus and tobacco mosaic virus), and plasmid expression vectors (eg, Ti plasmid) are suitable. Such cells are available from a variety of sources (eg, the American Type Culture Collection (ATCC), Manassas, VA; eg, Ausubel et al., “Current Protocols in Molecular Biology”, John). See also Wiley & Sons, New York, 1994). The method of transformation and the choice of expression vehicle will depend on the host system chosen. Transformation methods are described, for example, in Ausubel, supra. Expression vehicles are described, for example, in Pouwels et al., "Cloning Vectors: A Laboratory Manual", 1985, Supp. 1987.
[0042]
Host cells with an expression vehicle can be cultured in conventional nutrient media adapted as needed for activation or suppression of the selection gene, selection of transformants, or amplification of the selection gene.
[0043]
Where appropriate or beneficial, the nucleic acid encoding the fusion protein can include a signal sequence to secrete the fusion protein into cell culture and facilitate separation of the protein from the culture. Certain initiation signals may be required for efficient translation of the inserted nucleic acid sequence. Such signals include the ATG start codon and adjacent sequences. In some cases, an exogenous translational control signal, preferably including the ATG start codon, must be provided. Also, the start codon must be compatible with the reading frame of the desired coding sequence to ensure translation of the entire insertion. Such exogenous translational control signals and initiation codons are available from a variety of sources regardless of whether they are natural or synthetic. Efficiency of expression can be enhanced by the inclusion of appropriate transcriptional or translational enhancer sequences (eg, Bittner et al., Methods in Enzymol.153: 516, 1987).
[0044]
Component (1), component (2), and the fusion protein containing them can be solubilized under physiologically normal conditions. Such fusion proteins can also include at least a three-part fusion protein by including all or part of one or more unrelated proteins (ie, non-hsp, non-HPV proteins). . The “third” protein can be a protein that facilitates purification, detection, or solubilization of the fusion protein, or a protein that provides some other function. For example, the expression vector pUR278 (Ruther et al., EMBO J. Clin.2: 1791, 1983), and a lacZ fusion protein can be prepared, and a foreign polypeptide can be expressed as a fusion protein containing glutathione S-transferase (GST) using a pGEX vector. Generally, such fusion proteins are soluble and can be easily purified from lysed cells by adsorption to glutathione-agarose beads followed by elution in the presence of free glutathione. The pGEX vector is designed to include a thrombin or factor Xa protease cleavage site to cut off the cloned target gene product from the GST moiety. The "third" protein may be the Fc domain of an immunoglobulin. Such a fusion protein can be easily purified on an affinity column. It goes without saying that the fusion protein used in the method of the present invention comprises a plurality of components (1) and / or a plurality of components (2), and the components (1) and (2) are directly or indirectly linked. (Eg, linking such that one or more amino acid residues are present between the two).
[0045]
Proteins (eg, hsps, HPV antigens, or fusion proteins containing hsps) can be purified using antibodies that specifically bind the protein of interest. One skilled in the art can purify proteins using affinity-based purification methods. Methods for purifying non-denatured fusion proteins expressed in human cell lines are described, for example, in Janknecht et al., Proc. Natl. Acad. Sci. USA.88: 8972, 1981. In this system, the gene of interest is subcloned into a vaccinia recombination plasmid such that the open reading frame of the gene is translationally fused to an amino-terminal tag containing six histidine residues. Extract of cells infected with recombinant vaccinia virus2+Apply to a nitriloacetic acid-agarose column and selectively elute histidine-tagged proteins with a buffer containing imidazole. This same approach can be used for bacterial cultures.
[0046]
Proteins containing fusion proteins (especially those containing short antigenic fragments) were described by chemical synthesis (eg, “Solid Phase Peptide Synthesis”, Second Edition, 1984, The Pierce Chemical Co., Rockford, IL). Method).
[0047]
The isolated protein is further purified and / or concentrated, if desired, so long as further processing does not impair the ability to efficiently and sufficiently induce an immune response in the methods of the invention. Various methods for purifying and concentrating proteins are well known in the art (see, eg, Fisher, "Laboratory Technologies In Biochemistry And Molecular Biology", Ed. Work and Burdon, 1980, Elvis). See). Such methods include, for example, ultracentrifugation, and / or precipitation (eg, using ammonium sulfate), microfiltration (eg, using a 0.45 μm cellulose acetate filter), ultrafiltration (eg, sizing). Methods using membranes and recirculation filtration), gel filtration (e.g. Sepharose CL-6B, CL-4B, CL-2B, 6B, 4B or 2B, Sephacryl S-400 or S-300, Superose 6 or Ultrogel A2, Columns packed with A4 or A6; both available from Pharmacia), high performance protein liquid chromatography (FPLC), and high performance liquid chromatography (HPLC).
[0048]
Cross reactivity HPV Array
One of skill in the art can determine whether a composition comprising a first type of HPV antigen can be used to treat a subject infected with a second type of HPV. Assays on which such a determination can be made include predictive assays (eg, using computer models) and biological assays (eg, methods for examining cross-reactivity). One or both types of assays can be used (it is common practice to further study the results obtained in predictive assays in biological assays). Hereinafter, each example will be described.
[0049]
Cross-reactivity (ie, the ability of a composition comprising one type of HPV antigen to efficiently treat a subject who has become infected with another type of HPV or has a disease or condition associated with another type of HPV) ) Can be tested in well-established immunological methods. For example, double-transgenic (bi-transgenic) mice (Lustgarten et al., Human Immunol.) Generated to express the antigen-binding region of a human MHC class I molecule and the human CD8 gene.52: 109, 1997; Vitiello et al. Exp. Mod.173: 1007, 1991).
[0050]
More specifically, using HLA-A2 / CD8 double transgenic mice (Lustgarten et al., Supra), peptides derived from the HPV6 and 11 E7 proteins of cytotoxic T lymphocytes (CTL) against HPV16 E7. Cross-reactivity can be demonstrated by standard immunological techniques (see, eg, Colligan et al., “Current Protocols in Immunology”, John Wiley & Sons, 1999). Briefly, mice are immunized 1-3 times with an HspE7 fusion protein (based on BCG Hsp65 and HPV16 E7 molecules) at an interval of 7-21 days. HspE7 is suspended in phosphate buffered saline (PBS) and administered subcutaneously in a dose range of 1 μg to 1000 μg per animal. Seven days after the last HspE7 administration, the mice are sacrificed, the spleen is removed, and the tissue is broken up into a single cell suspension. The CTL specific for HPV E7 is restimulated by adding the HLA-A2 binding peptide derived from HPV16 E7, HPV6 E7, and HPV11 E7 to the medium at a concentration of 1 μM. The cells are restimulated, for example, in a 6 well plate with a different peptide in each well. The peptides predicted by computer algorithms to have the highest HLA-A2 binding affinity (eg, 10 peptides) were identified as HPV16, HPV6, and HPV11 (or any other HPV type; Parker et al., J. Immunol.152163, 1994; such an algorithm is available from the National Institutes of Health (NIH) 's BIMAS (Bioinformatics & Molecular Analysis Section) website (http: //bimas.dcrtt as of June 26, 2001). .Nih.gov /)). In still other cases, the corresponding peptides from the other two HPV genotypes (peptides 11-20 of HPV16 E7 and peptides 11-20 of HPV6 and 11) can also be used.
[0051]
After a period of time (eg, one week) after restimulation in vitro, CTL activity is measured by lysing T2 target cells treated with HLA-A2-binding peptides derived from HPV16 E7, HPV6 E7, and HPV11 E7. be able to. In addition, antigen-specific T lymphocytes recognizing HLA-A2 binding peptides derived from HPV16 E7, HPV6 E7, and HPV11 E7 can be measured by ELISPOT analysis of IFN-γ-secreting cells according to literature methods (Asal Et al., Clin. Diagn. Lab. Immunol.7145, 2000). This assay can be performed on transgenic mice of other HLA alleles.
[0052]
Alternatively, the ability of CTLs induced by HspE7 immunization to cross-react with peptides derived from the E7 protein of HPV6 or 11 in a human HLA-A2-positive subject undergoing treatment of genital warts with HspE7 Can be measured. Prior to treatment, and several days (eg, 7 days) after each treatment with HspE7, peripheral blood mononuclear cells (PBMCs) can be isolated from a subject (eg, a human patient). Such cells contain a fluorogenic MHC-peptide complex (tetramer, Altman et al., Science).274: 94, 1996) or by ELISPOT analysis (Asal et al., Clin. Diagn. Lab. Immunol.7145, 2000). Cells were assayed directly from peripheral blood and analyzed by Youde et al. (Cancer Res.60: 365, 2000) by in vitro restimulation. For in vitro restimulation, 2 × 106/ Ml of PBMC is cultured in RPMI 1640 with 10% human AB serum (RAB) and peptide (10 μg / ml). The restimulatory peptide may be derived from HPV16 E7 and include peptides predicted to have the highest HLA-A2 binding affinity (eg, 10 peptides) as defined by a computer algorithm (Parker et al., Supra). it can. On day 4, 1 ml RAB containing 25 units / ml IL-2 is added to each well. On day 6, replace 1 ml of medium with 1 ml of medium containing 10 units / ml of IL-2. On the 7th day, 3 × 10 3 irradiated autologous PBMCs (fresh or thawed after freezing)6 At a concentration of cells / ml, 10 μg / ml peptide and 3 μg / ml β2-Resuspend in RAB containing microglobulin. After allowing the antigen-presenting cells to adhere for 2 hours, the cells were washed to remove non-adherent cells, and 1-2 × 106 Add effector cells / ml. On day 9, 1 ml RAB containing 25 units / ml IL-2 is added to each well. On day 13, the contents of the wells are split into multiplates and medium (containing 10 units / ml of IL-2) is added to the original volume. Day 14 cells are used. For FACS analysis, tetramers are prepared by literature methods (Altman et al., Science 274: 94, 1996). The peptide used for the addition of the tetramer is an HLA-A2 binding peptide derived from the E7 molecule of HPV16, HPV6, and HPV11. The peptide (eg, 10 peptides) predicted to have the highest HLA-A2 binding affinity by a computer algorithm (Parker et al., Supra) is used for each of HPV16, HPV6, and HPV11. In still other cases, the corresponding peptides from the other two HPV genotypes (peptides 11-20 of HPV16 E7 and peptides 11-20 of HPV 6 and 11) are also used. Fresh or restimulated PBMCs are stained with PE-labeled HPV-E7 peptide tetramer and FITC-labeled anti-CD8 antibody and analyzed by flow cytometry according to literature procedures. ELISPOT analysis of antigen-specific T lymphocytes recognizing HLA-A2 binding peptides from HPV16 E7, HPV6 E7, and HPV11 E7 present in fresh and restimulated PBMCs has been described in the literature. (Asal et al., Clin. Diagn. Lab. Immunol,7145, 2000). Similarly, the above method can be applied to a subject having another HLA haplotype.
[0053]
Human PBMCs from HLA-A2-positive healthy subjects who have not been treated with HspE7 in the past and have not been stimulated in vitro with HspE7 proteins or peptides derived from HPV 16 type E7, The ability to cross-react with cells treated with corresponding peptides derived from HPV genotypes (types 6 and 11) can be examined. Stimulation and analysis of cells is performed using procedures common in the art. Briefly, PBMCs are isolated from peripheral blood, adherent cells are separated from non-adherent cells, adherent cells are cultured, and dendritic cells (DCs) are purified from “Current Protocols in Immunology” (“Current Protocols in Immunology”). Colligan et al., Edited by John Wiley & Sons, pp. 7.32.7-8, 1999). Non-adherent cells are stored frozen in 90% FCS / 10% DMSO and used at each time point in later assays.
[0054]
Upon stimulation, 50 μg / ml HspE7 or 40 μg / ml of the appropriate peptide and 3 μg / ml β2-5% CO at 37 ° C in microglobulin for 24 hours2Treat the DC under (Kawashima et al., Human Immunol.59: 1, 1998). The peptide used is an HLA-A2 binding peptide derived from the E7 molecule of HPV16, HPV6 and HPV11. The peptides predicted by computer algorithms (Parker et al., Supra) to have the highest HLA-2 binding affinity (eg, 10 peptides) are used for each of HPV16, HPV6, and HPV11. In other cases, the corresponding peptides from the other two HPV genotypes (peptides 11-20 of HPV16 E7 and peptides 11-20 of HPV 6 and 11) are also used. CD8+Cells are isolated from cryopreserved autologous non-adherent cells by positive selection using immunomagnetic beads (Miltenyi Biotec). The peptide / protein-loaded DCs were irradiated with 4200 rads of radiation and autologous CD8+A ratio of 1:20 with cells, eg 0.25 × 10 5 in 0.5 ml RAB5 DC and 5 × 105 CD8+Mix cells in a 48 well plate containing 10 ng / ml IL-7. On days 7 and 14, the cells are restimulated with adherent APC treated with the autologous peptide (Kawashima et al., Human Immunol.59: 1, 1998). Add 10 U / ml hIL-2 to the culture every 2-3 days. T lymphocytes specific for the HPV E7 peptide were elicited at 7 and 14 days after in vitro stimulation with a fluorogenic MHC-peptide complex (tetramer, Altman et al., Science).274: 94, 1996), or ELISPOT analysis (Asal et al., Clin. Diagn. Lab. Immimol.7145, 2000). For FACS analysis, tetramers were analyzed by literature methods (Altman et al., Science).274: 94, 1996). The peptide used for the addition of the tetramer can be an HLA-A2 binding peptide derived from the E7 molecule of HPV16, HPV6, and HPV11, as described above. T lymphocytes specific for the peptide are stained with PE-labeled HPV-E7 peptide tetramer and FITC-labeled anti-CD8 antibody and analyzed by flow cytometry (Youde et al., Cancer Res.60: 365, 2000). ELISPOT analysis of antigen-specific T lymphocytes recognizing HLA-A2 binding peptides derived from HPV16 E7, HPV6 E7, and HPV11 E7 was performed according to literature methods (Asal et al., Clin. Diagn. Lab. Immol.7145, 2000). Similarly, the above method can be applied to a subject having another HLA haplotype.
[0055]
Administration of the composition
The present invention relates to an HPV protein antigen (for example, an HPV protein antigen (eg, an HPV protein antigen (or an antigenic fragment thereof)), an HPV protein antigen mixed with or prepared with an hsp (or an immunostimulatory fragment thereof), Or an antigenic fragment thereof) and a fusion protein comprising hsp (or an immunostimulatory fragment thereof). Optionally, these proteins can be diluted (e.g., PBS) or bicarbonate solution (e.g., 0.24 M NaHCO3).3) Can be suspended in a pharmaceutically acceptable carrier. Useful carriers can be selected based on the mode and route of administration and by standard pharmaceutical practice. Suitable pharmaceutical carriers and diluents, as well as pharmaceutically necessary substances for use, are described in "Remington's Pharmaceutical Sciences". Adjuvants such as cholera toxin, E. coli heat labile enterotoxin (LT), liposomes, or immunostimulatory complexes (ISCOM) can also be included.
[0056]
It is not necessary to administer the protein (eg, a fusion protein) directly to the subject; instead, a nucleic acid sequence encoding the protein can be administered. Such proteins are expressed in vivo in a subject. The nucleic acid can be part of a vector (such as a viral vector, such as a viral vector genome), or can be, for example, encapsulated in a liposome. Nucleic acids can also be administered as naked nucleic acids.
[0057]
The composition can be formulated as a solution, suspension, suppository, tablet, granule, powder, capsule, ointment, or cream. As noted above, one or more pharmaceutical carriers can be included in preparing the compositions. Examples of pharmaceutically acceptable carriers or other additives include solvents (eg, water or saline), solubilizing agents (eg, ethanol, polysorbate, or Cremophor EL®), to adjust isotonicity. Drugs, preservatives, antioxidants, excipients (eg, lactose, starch, microcrystalline cellulose, mannitol, maltose, calcium phosphate, light anhydrous silicic acid, or calcium carbonate), binders (eg, starch, polyvinylpyrrolidone, Hydroxypropylcellulose, ethylcellulose, carboxymethylcellulose or gum arabic), a lubricant (eg, magnesium stearate, talc, or hardened oil), or a stabilizer (eg, lactose, mannitol, maltose, polysorbate, macrogel, or polyoxyethylene hard) Castor oil), and the like. If necessary (or desirable), glycerin, dimethylacetamide, sodium lactate, surfactant, sodium hydroxide, ethylenediamine, ethanolamine, sodium carbonate, arginine, meglumine, or trisaminomethane can be used. . Biodegradable polymers such as poly-D, L-lactide-co-glycolide or polyglycolide can be used as the bulk matrix if sustained release of the composition is desired (eg, US Pat. (See Patents 5,417,986, 4,675,381, and 4,450,150). Pharmaceutical preparations, such as solutions, tablets, granules, or capsules, can be made using the above compositions. When the composition is administered orally, flavoring and coloring agents can be added.
[0058]
Therapeutic compositions may be administered by any suitable route, e.g., intravenously, intraarterially, topically, intraperitoneally, intrapleurally, orally, subcutaneously, intramuscularly, intradermally, sublingually, subepidermally, intranasally, pulmonary. It can be administered internally (eg, by inhalation), vaginally, or rectally.
[0059]
The amount of the composition to be administered depends, for example, on the particular composition, whether the adjuvant is co-administered with the composition, the type of adjuvant co-administered, the mode and frequency of administration, and the desired effect (eg, prevention or treatment). ). Dosages are determined by those of ordinary skill in the art during the course of drug or prophylactic drug development. Generally, the compositions of the present invention will be administered in dosages from 1 μg to 100 mg for adult humans. When an adjuvant is administered with the composition, an amount of 1 ng to 1 mg per adult human can generally be used. Administration is repeated as needed as determined by one skilled in the art. For example, three booster doses can be given once a week or once a month after the first dose. Booster administration can be performed at 3 to 12 weeks after the first administration. The second booster administration can be performed after 3 to 12 weeks using the same dosage form. Serum or T cells can be obtained from a subject and used, for example, to study the immune response generated by the composition against the fusion protein or HPV antigens contained in the protein conjugate. Methods for examining antibodies or cytotoxic T cells for a particular antigen are well known in the art. Additional booster administrations can be performed as needed. For example, by varying the amount of fusion protein included in the composition, the immunization protocol can be optimized to elicit a maximal immune response.
[0060]
It will be appreciated that the proteins described herein can be administered using nucleic acids such as viral vectors (eg, retroviral or adenoviral vectors).
[0061]
Without further elaboration, it is believed that one skilled in the art can, using the preceding description and the following examples, utilize the present invention to its fullest extent. The following examples are to be construed merely to illustrate that those skilled in the art can isolate and use the fusion polypeptides, and do not limit the remainder of the disclosure in any way.
[0062]
Example
A fusion polypeptide containing Mycobacterium bovis BCG Hsp65 to which HPV type 16 E7 protein was bound was recombinantly produced and formulated by the procedure described in WO 99/07860. Hsp65 is a molecule of the Hsp60 family of stress proteins. The following observations were made during the course of human clinical trials examining the efficacy of this fusion polypeptide in treating severe squamous intraepithelial lesions of the anus (HSIL).
[0063]
Twenty-two patients participated in a randomized, double-blind, placebo-controlled, multicenter study of HspE7 in anal HSIL treatment. Eligible patients were confirmed by biopsy to have anal HSIL and had a negative human immunodeficiency virus (HIV) test result. The patient's HPV type was determined using cells obtained from an anal examination and did not need to have HPV-16. The HPV type of individual lesions was not determined. Patients received three subcutaneous injections of 100 μg HpsE7 or placebo each month. Treatment response was also assessed by anal papilloma colony testing, high-resolution anoscopy with biopsy (HRA), and physician-wide evaluation. Non-responders (who have persistent anal HSIL) after 12 or 24 weeks in the control study were switched to open-label studies. In the open-label study, patients received three monthly injections of 500 μg of HspE7. The evaluation of treatment was double-blind in a placebo-controlled trial, and no treatment was disclosed.
[0064]
Using Dacron swabs to determine the patient's HPV type, samples were taken from the patient's anus immediately prior to biopsy at screening at randomized placebo-controlled trials. DNA was isolated after transfer to Sample Transport Medium (Digene) and used for HPV type determination. Briefly, HPV DNA was amplified by polymerase chain reaction (PCR) using consensus primer set MY09 / MY11. Following the amplification step, samples were blotted onto nylon membranes and 29 HPV types
[0065]
To verify the success of the PCR, a β-globin control was amplified and probed for each sample. If the sample did not test positive for the presence of β-globin, the PCR step was considered technically failed. A control sample was considered "HPV negative" if the consensus probe did not give a score of two or more.
[0066]
At the time of entry to the open-label study, 14 out of 22 patients (64%) received sustained genital warts through a prior double-blind study in which 100 μg of HspE7 or placebo were injected three times monthly. Luxury was acknowledged. Of these 14 patients, 8 (57%) had worsened, 4 (29%) did not change, and 2 (14%) improved (one dramatically improved and another slightly improved). Another patient had condyloma at the beginning of the double-blind study, but resolved before the open-label study began, along with the other seven who did not detect warts during any study. Excluded from analysis. Condyloma was found inside the anorectal tract in all 14 patients (100%) and in the perianal skin in 6 of 14 (43%). Investigators reported that of 14 patients with warts present at the start of the open-label study, 11 patients (79%) who had surgical resection had local excision (eg, liquid nitrogen or electrocautery) ) Was required for two patients (14%) and topical therapy (imiquimod) was required for one patient (7%), respectively. These patients chose to delay treatment recommended by the investigator and instead agreed to receive three monthly injections of 500 μg HspE7 in an open-label study.
[0067]
One month after the last treatment with 500 μg of HspE7, 2 patients (14%) had no detectable warts and 11 patients (79%) There was a decrease in wart size or number compared to the situation, and one patient (7%) had an increase in wart size (Table 1). By the time of the initial evaluation of the open-label study (four months after the last dose), one more patient had an effective to significant improvement (no visible warts). For this reason, a total of three (21%) showed a remarkable effect (Table 1). None of these responders relapsed during the 6-month evaluation period of the open-label study. Ten patients (71%) continued to show effective improvement (ie, the wart size was further significantly reduced, and the degree of treatment needed to remove the remaining wart was continuously reduced) did). One non-responder (7%) did not improve by the end of the open-label study
[0068]
Table 1: List of responses of genital warts after treatment with HspE7
4 months after final treatment with 500 μg HspE7
[0069]
At the end of the study, the investigator did not recommend continuing treatment for the three responders. As shown in Table 2, the treatment recommended by the investigator for those with effective results included ablation therapy (6 of 14; 43%) or treatment with topical drugs (14 (4/29%). Non-responders were recommended for additional surgery (1 in 14; 7%). All 22 agreed to proceed with the registry protocol for long-term follow-up of the response and postpone the treatment recommended by the attending physician.
[0070]
Table 2: Evaluation of anal genital response and treatment recommended by physician
One month after the last treatment with 500 μg of HspE7
4 months after final treatment with 500 μg HspE7
Abbreviation: CR = Effective; PR = Effective
[0071]
In all 14 individuals diagnosed with genital warts, HPV DNA of multiple HPV types was detected in anal smears during the screening of the first randomized controlled trial (Table 3). HPV-6 and / or 11 were found in 12 patients (86%). In one patient, only HPV-16 and related types were observed; in other patients, the type could not be determined. Three out of 14 patients (21%) were HPV-16 positive. Most of the patients with improved warts (11/13, 85%) did not have HPV-16. No HPV-16 was observed in non-responders (see Table 3).
[0072]
[Table 3] HPV type of patients with genital warts
4 months after final treatment with 500 μg HspE7
Abbreviation: HPV = human papillomavirus; CR = effective; PR = effective
[0073]
An open-label crossover study in patients with persistent anal HSDL and concomitant anogenital warts receiving HspE7 (500 μg q 3 months) showed warts at baseline 14 In three of the patients (17%), no warts were found four months after the last treatment. Another 10 patients (71%) had improved symptoms (the wart size was significantly reduced, and the level of treatment needed to remove the remaining wart continued to decrease) . In one patient (7%), no improvement was noted during the study, and additional surgery was recommended by the investigator.
[0074]
Prior to enrollment in the open-label study, most patients at the trial site had undergone surgical intervention for wart removal (11 of 14; 79%). By the end of the study, only one was recommended for surgical treatment. Local ablation therapy (eg, liquid nitrogen, electrocautery) was recommended for 6 patients (43%), and treatment with topical drugs (eg, imiquimod) was recommended for 4 patients (29%). Three patients did not require subsequent treatment.
[0075]
The response at 6 months was considered progressive and no responders recurred during the same period. The progressive and progressive resolution of condyloma does not differ from predictions based on immunological host responses after induction of cellular immunity by HspE7.
[0076]
Two patients who participated in the double-blind study had some improvement in condyloma before proceeding to the open-label study. At this time we have not been informed of the nature of the randomization and it is unclear whether these patients received 100 μg of HspE7 or a placebo. However, based on the response observed in an open-label test with three monthly injections with 500 μg of HspE7, the higher dose is likely to be more active than at 100 μg.
[0077]
HPV-16 DNA was detected in anal smears in only 3 out of 13 patients (23%) who had improved warts after treatment with HspE7. DNA from HPV-6, HPV-11, or both, was detected in the majority of patients who responded to warts with treatment with HspE7. The above data suggest that there is immunological cross-reactivity between the HPV types in these responses to HspE7.
[0078]
In summary, the results presented herein suggest that HspE7 is widely active against genital warts. This activity does not appear to be restricted to HPV-16 positive patients but crosses multiple HPV types. HspE7 has shown activity in the treatment of genital diseases caused by HPV, and this activity is not thought to be restricted to HPV-16 positive patients.
[0079]
The observations reported here suggest that treatment with HspE7 may be a new, simple and non-surgical treatment for genital warts. And, at least, by reducing the wart load, it may reduce the degree of treatment and the resulting morbidity. Disorders within the anorectal may require additional treatment of debilitating, debilitating pain. Treatment that reduces or eliminates the amount or degree of "surgical" or ablation therapy, resulting in a partial response, also reduces morbidity, shortens working time, and improves QOL.
[0080]
The above results indicate that the heat shock protein / HPV type 16 antigen composition is effective in removing or reducing warts predominantly due to HPV types 6 and 11. It is important to note that (1) warts can be treated with all HPV-based compositions, and (2) HPV 16-type compositions can be used to treat conditions thought to be due to HPV other than type 16. The result is that it is effective. The result of the latter cross-reactivity was almost unpredictable given the general understanding that type-specific compositions produce only type-specific immune responses.
[0081]
To investigate the mechanism of cross-reactivity observed for the fusion polypeptide, theoretical binding was calculated for various HLA class I molecules and HPV types 16, 6 and 11 E7 peptides. Dissociation T1/2Is described by Parker et al. Immunol.152163, 1994 (see also the website mentioned above). The data is summarized in Table 4.
[0082]
[Table 4]
[0083]
The results in Table 4 suggest that, depending on the particular HLA molecule under investigation, HPV 16 type E7 antigens can elicit a cellular immune response against other HPV type E7 antigens. For example, in the case of HLA B 2705, the presence of high levels of binding was predicted for peptides starting at amino acid position 76 of E7 of all three HPV types. Thus, in patients expressing the same HLA molecule, HPV 16 type E7 compositions are likely to cross-react and be useful in treating or preventing HPV type 6 and 11 infections. Each peptide fragment listed in bold in Table 4 is an antigenic fragment that can be included in a composition (eg, the fusion polypeptide described herein) as a substrate for the complete E7 virus antigen. It goes without saying that longer fragments containing two or more putative HLA epitopes, or many putative HLA epitopes, can also be used.
Claims (39)
（１）ｈｓｐまたはその免疫刺激性断片、および（２）ＨＰＶのタンパク質またはその抗原性断片を含む、融合タンパク質を含む組成物を対象に投与する段階であって、該対象が、対象に投与されるＨＰＶ型とは異なるＨＰＶ型に感染していて、疾患または症状を治療するのに十分な量の組成物が投与される段階。A method of treating a subject exhibiting a human papillomavirus (HPV) -related disease or condition in the subject, comprising:
Administering to the subject a composition comprising a fusion protein comprising (1) hsp or an immunostimulatory fragment thereof, and (2) a protein of HPV or an antigenic fragment thereof, wherein the subject is administered to the subject. Infecting a different HPV type than the HPV type, and administering a sufficient amount of the composition to treat the disease or condition.
疣贅を有する対象、または疣贅を有することが疑われる対象を同定する段階；
（１）ｈｓｐまたはその免疫刺激性断片、および（２）ＨＰＶのタンパク質、またはその抗原性断片を含む融合ポリペプチドをコードする核酸を対象に投与する段階；ならびに
疣贅を治療するのに十分な量の融合ポリペプチドを対象で発現させる段階。How to treat a wart in a subject, including the following steps:
Identifying a subject having, or suspected of having, a wart;
(1) administering to the subject a nucleic acid encoding a fusion polypeptide comprising hsp or an immunostimulatory fragment thereof, and (2) a protein of HPV, or an antigenic fragment thereof; and sufficient to treat warts. Expressing an amount of the fusion polypeptide in the subject.
（１）ｈｓｐまたはその免疫刺激性断片、および（２）ＨＰＶのタンパク質を含む、融合タンパク質をコードする核酸を対象に投与する段階であって、対象に投与されるＨＰＶ型とは異なるＨＰＶ型に対象が感染している段階、ならびに
疾患または症状を治療するのに十分な量の融合タンパク質を、対象において発現させる段階。A method of treating a subject having a disease or condition associated with an HPV infection, comprising the following steps:
Administering to the subject a nucleic acid encoding a fusion protein comprising (1) hsp or an immunostimulatory fragment thereof, and (2) a HPV protein, wherein the HPV type is different from the HPV type administered to the subject. The stage at which the subject is infected, and the stage at which the fusion protein is expressed in the subject in an amount sufficient to treat the disease or condition.
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| PCT/US2001/020240 WO2002000242A2 (en) | 2000-06-26 | 2001-06-26 | Human papilloma virus treatment |
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| CA2378097A1 (en) | 1999-07-08 | 2001-01-18 | Stressgen Biotechnologies Corporation | Induction of a th1-like response in vitro |
| US20050226890A1 (en) * | 1999-08-12 | 2005-10-13 | Cohen David I | Tat-based vaccine compositions and methods of making and using same |
| US20050100928A1 (en) * | 1999-09-16 | 2005-05-12 | Zycos Inc., A Delaware Corporation | Nucleic acids encoding polyepitope polypeptides |
| US8128922B2 (en) | 1999-10-20 | 2012-03-06 | Johns Hopkins University | Superior molecular vaccine linking the translocation domain of a bacterial toxin to an antigen |
| NZ523408A (en) * | 2000-06-26 | 2006-02-24 | Stressgen Biotechnologies Corp | Human papilloma virus treatment |
| CA2417214C (en) | 2000-08-03 | 2016-06-21 | Johns Hopkins University | Molecular vaccine linking an endoplasmic reticulum chaperone polypeptide to an antigen |
| WO2005090392A1 (en) * | 2004-03-16 | 2005-09-29 | Inist Inc. | Tat-based tolerogen compositions and methods of making and using same |
| RU2295536C2 (en) | 2001-02-05 | 2007-03-20 | Стрессджен Байотекнолоджиз Корп. | Composition based on hepatitis b virus (hbv) protein and stress protein and uses thereof |
| US6649402B2 (en) * | 2001-06-22 | 2003-11-18 | Wisconsin Alumni Research Foundation | Microfabricated microbial growth assay method and apparatus |
| CA2502696A1 (en) * | 2002-10-21 | 2004-05-06 | Mgi Pharma Biologics, Inc. | Compositions and methods for treating human papillomavirus-mediated disease |
| KR20050086924A (en) * | 2002-12-20 | 2005-08-30 | 글락소스미스클라인 바이오로지칼즈 에스.에이. | Hpv-16 and -18 l1 vlp vaccine |
| WO2004098526A2 (en) | 2003-05-05 | 2004-11-18 | Johns Hopkins University | Anti-cancer dna vaccine employing plasmids encoding signal sequence, mutant oncoprotein antigen, and heat shock protein |
| WO2005026375A2 (en) | 2003-05-22 | 2005-03-24 | Fraunhofer Usa, Inc. | Recombinant carrier molecule for expression, delivery and purification of target polypeptides |
| DE10347710B4 (en) * | 2003-10-14 | 2006-03-30 | Johannes-Gutenberg-Universität Mainz | Recombinant vaccines and their use |
| US7285386B2 (en) * | 2003-10-29 | 2007-10-23 | University Of New Mexico, And Educational Institution Of The State Of New Mexico | RhPV as a model for HPV-induced cancers |
| DE10357677A1 (en) * | 2003-12-10 | 2005-07-14 | Greiner Bio-One Gmbh | Primers and probes for detecting genital HPV genotypes |
| US7927580B2 (en) * | 2004-03-16 | 2011-04-19 | Nanirx, Inc. | Tat-based immunomodulatory compositions and methods of their discovery and use |
| WO2006073970A2 (en) | 2005-01-06 | 2006-07-13 | The Johns Hopkins University | Rna interference that blocks expression of pro-apoptotic proteins potentiates immunity induced by dna and transfected dendritic cell vaccines |
| US8252893B2 (en) * | 2005-01-31 | 2012-08-28 | Board Of Trustees Of The University Of Arkansas | CD8 T cell epitopes in HPV 16 E6 and E7 proteins and uses thereof |
| ATE506063T1 (en) * | 2005-04-27 | 2011-05-15 | Univ Leiden Medical Ct | TREATMENT OF HPV-INDUCED INTRAEPITHELIAL ANOGENITAL NEOPLASIA |
| CA2616859C (en) * | 2005-08-03 | 2015-04-14 | Fraunhofer Usa, Inc. | Compositions and methods for production of immunoglobulins |
| US7622121B2 (en) * | 2005-09-21 | 2009-11-24 | New York University | Heat shock proteins from Mycobacterium leprae and uses thereof |
| US20080279877A1 (en) * | 2006-02-13 | 2008-11-13 | Fraunhofer U.S.A. Inc. | HPV antigens, vaccine compositions, and related methods |
| JP2009526526A (en) * | 2006-02-13 | 2009-07-23 | フラウンホーファー ユーエスエー， インコーポレイテッド | Influenza antigens, vaccine compositions, and related methods |
| US8277816B2 (en) * | 2006-02-13 | 2012-10-02 | Fraunhofer Usa, Inc. | Bacillus anthracis antigens, vaccine compositions, and related methods |
| CA2653474A1 (en) * | 2006-05-31 | 2007-12-06 | Nventa Biopharmaceuticals Corporation | Bioactive purified hspe7 compositions |
| US9085638B2 (en) | 2007-03-07 | 2015-07-21 | The Johns Hopkins University | DNA vaccine enhancement with MHC class II activators |
| WO2008124483A1 (en) * | 2007-04-04 | 2008-10-16 | Specigen, Inc. | Protein cage immunotherapeutics |
| EP2152301A4 (en) * | 2007-04-28 | 2010-07-28 | Fraunhofer Usa Inc | Trypanosoma antigens, vaccine compositions, and related methods |
| US8404252B2 (en) | 2007-07-11 | 2013-03-26 | Fraunhofer Usa, Inc. | Yersinia pestis antigens, vaccine compositions, and related methods |
| US20110059130A1 (en) * | 2007-08-20 | 2011-03-10 | Fraunhofer Usa, Inc. | Prophylactic and therapeutic influenza vaccines, antigens, compositions and methods |
| US20090117140A1 (en) * | 2007-09-26 | 2009-05-07 | Mayumi Nakagawa | Human papilloma virus dominant CD4 T cell epitopes and uses thereof |
| WO2009094006A2 (en) * | 2007-10-25 | 2009-07-30 | Wake Forest University Health Sciences | Bordetella outer-membrane protein antigens and methods of making and using the same |
| AU2009223838B2 (en) | 2008-03-03 | 2012-07-26 | The University Of Miami | Allogeneic cancer cell-based immunotherapy |
| CA2718884C (en) | 2008-03-20 | 2016-11-22 | University Of Miami | Heat shock protein gp96 vaccination and methods of using same |
| WO2010037046A1 (en) | 2008-09-28 | 2010-04-01 | Fraunhofer Usa, Inc. | Humanized neuraminidase antibody and methods of use thereof |
| AU2009309616B2 (en) * | 2008-10-31 | 2014-02-13 | Novartis Ag | Combination of a phosphatidylinositol-3-kinase (PI3K) inhibitor and a mTOR inhibitor |
| CN102227221A (en) * | 2008-11-28 | 2011-10-26 | 诺瓦提斯公司 | Pharmaceutical combination containing hsp90 inhibitor and mtor inhibitor |
| CN102405057B (en) | 2009-03-23 | 2016-05-25 | 那尼尔科斯治疗公司 | By immunostimulating Hiv Tat derivative polypeptides treatment cancer |
| WO2011041391A1 (en) | 2009-09-29 | 2011-04-07 | Fraunhofer Usa, Inc. | Influenza hemagglutinin antibodies, compositions, and related methods |
| US11963716B2 (en) | 2010-07-19 | 2024-04-23 | Emblation Limited | Apparatus and method for the treatment of dermatological diseases or conditions |
| US9662510B2 (en) | 2010-07-19 | 2017-05-30 | Emblation Limited | Apparatus and method for the treatment of dermatological diseases or conditions |
| KR20140045341A (en) * | 2011-02-23 | 2014-04-16 | 유니버시티 오브 마이애미 | Combined cell based gp96-ig-siv/hiv, recombinant gp120 protein vaccination for protection from siv/hiv |
| US9028884B2 (en) | 2013-08-13 | 2015-05-12 | Preventamedics LLC | Medical delivery devices and methods for applying a barrier composition to a targeted skin surface |
| WO2015051245A1 (en) | 2013-10-04 | 2015-04-09 | Pin Pharma, Inc. | Immunostimulatory hiv tat derivative polypeptides for use in cancer treatment |
| GB2541749B (en) | 2015-08-31 | 2020-12-09 | Emblation Ltd | An interference suppression apparatus and method |
| US11497926B2 (en) | 2016-08-08 | 2022-11-15 | Emblation Limited | Method and apparatus for the treatment, management and/or control of pain |
| US20230241207A1 (en) * | 2017-04-03 | 2023-08-03 | Neon Therapeutics, Inc. | Protein antigens and uses thereof |
| CN108409866A (en) * | 2018-01-23 | 2018-08-17 | 合肥瑞城生生物科技有限公司 | A kind of multi-epitope combined peptide for treating and preventing human papilloma virus infection and relevant disease |
| KR20210044805A (en) | 2018-08-06 | 2021-04-23 | 닐슨 바이오사이언스, 아이엔씨. | Wart treatment |
| CN114478712B (en) * | 2022-03-29 | 2022-09-23 | 深圳吉诺因生物科技有限公司 | HPV epitope and identification method and application thereof |
Family Cites Families (73)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4666847A (en) * | 1981-01-16 | 1987-05-19 | Collaborative Research, Inc. | Recombinant DNA means and method |
| NO172595C (en) | 1983-02-07 | 1993-08-11 | Battelle Memorial Institute | PROCEDURE AND EXPRESSION PLASMID FOR THE PREPARATION OF HEMAGGLUTININ UNDER CONTROL OF DROSOPHILA HSP PROMOTER |
| US4797359A (en) * | 1983-05-10 | 1989-01-10 | Board Of Regents, The University Of Texas System | Heat shock regulated production of selected and fused proteins in yeast |
| GB8404280D0 (en) | 1984-02-17 | 1984-03-21 | Stanford J L | Biological preparations |
| IL71683A0 (en) | 1984-04-27 | 1984-09-30 | Yeda Res & Dev | Pharmaceutical compositions for treating arthritis type diseases comprising fractions obtained from mycobacteria |
| US4918164A (en) * | 1987-09-10 | 1990-04-17 | Oncogen | Tumor immunotherapy using anti-idiotypic antibodies |
| US4784941A (en) * | 1985-04-08 | 1988-11-15 | Genetic Systems Corporation | Expression and diagnostic use of pENV-3 encoded peptides which are immunologically reactive with antibodies to LAV |
| JPS62164696A (en) | 1986-01-06 | 1987-07-21 | エフ．ホフマン ― ラ ロシュ アーゲー | Development of htlv-iiigag gene |
| US4734362A (en) * | 1986-02-03 | 1988-03-29 | Cambridge Bioscience Corporation | Process for purifying recombinant proteins, and products thereof |
| US4906742A (en) | 1986-07-31 | 1990-03-06 | Whitehead Institute For Biomedical Research | Encoding antigens of M. Leprae |
| US6007806A (en) | 1986-08-13 | 1999-12-28 | Transgene S.A. | Expression of a tumor-specific antigen by a recombinant vector virus and use thereof in preventive or curative treatment of the corresponding tumor |
| NL8701163A (en) | 1986-09-09 | 1988-04-05 | Nederlanden Staat | USE OF A PEPTIDE FOR THE PREPARATION OF PREPARATIONS FOR LIGHTING, TREATMENT AND DIAGNOSIS OF AUTOIMMUNE DISEASES, IN PARTICULAR ARTHRITIC CONDITIONS, COMPOUNDS RELATED TO THIS PEPTIDE, AND PHARMACEUTICAL AND DIAGNOSTIC PREPARATORY. |
| AU1548388A (en) | 1987-02-02 | 1988-08-24 | Whitehead Institute For Biomedical Research | Mycobacterium tuberculosis genes and encoding protein antigens |
| US4952395A (en) | 1987-02-26 | 1990-08-28 | Scripps Clinic And Research Foundation | Mycobacterial recombinants and peptides |
| US5504005A (en) | 1987-03-02 | 1996-04-02 | Albert Einstein College Of Medicine Of Yeshiva University | Recombinant mycobacterial vaccine |
| US4918166A (en) | 1987-04-10 | 1990-04-17 | Oxford Gene Systems Limited | Particulate hybrid HIV antigens |
| US5256767A (en) * | 1987-06-10 | 1993-10-26 | The Immune Response Corporation | Retroviral antigens |
| NL8703107A (en) | 1987-12-22 | 1989-07-17 | Nederlanden Staat | POLYPEPTIDES AND DERIVATIVES THEREOF, AND THEIR USE THEREOF IN PHARMACEUTICAL AND DIAGNOSTIC PREPARATIONS. |
| US5204259A (en) * | 1988-05-06 | 1993-04-20 | Pharmacia Genetic Engineering, Inc. | Methods and systems for producing HIV antigens |
| WO1989012455A1 (en) | 1988-06-15 | 1989-12-28 | Whitehead Institute For Biomedical Research | Stress proteins and uses therefor |
| WO1990010230A1 (en) | 1989-02-23 | 1990-09-07 | University Of Ottawa | Polypeptide having immunological activity for use as diagnostic reagent and/or vaccine |
| US5114844A (en) | 1989-03-14 | 1992-05-19 | Yeda Research And Development Co., Ltd. | Diagnosis and treatment of insulin dependent diabetes mellitus |
| JPH04506297A (en) | 1989-06-19 | 1992-11-05 | ホワイトヘツド・インスチチユート・フオー・バイオメデイカル・リサーチ | Vector-mediated genomic insertion and expression of DNA in BCG |
| GB8919321D0 (en) | 1989-08-25 | 1989-10-11 | Univ London | Treatment of chronic inflammatory conditions |
| GB9007194D0 (en) | 1990-03-30 | 1990-05-30 | Wellcome Found | Live vaccines |
| US5348945A (en) | 1990-04-06 | 1994-09-20 | Wake Forest University | Method of treatment with hsp70 |
| AU660430B2 (en) | 1990-11-08 | 1995-06-29 | Stanford Rook Limited | Mycobacterium as adjuvant for antigens |
| GB9024320D0 (en) | 1990-11-08 | 1990-12-19 | Univ London | Treatment of uveitis |
| GB2251186A (en) | 1990-12-04 | 1992-07-01 | Randall Neal Gatz | Polypeptide for use in treatment of autoimmune disease |
| EP0521220A1 (en) | 1991-06-14 | 1993-01-07 | Institut Pasteur | Recombinant immunogenic actinomycetale |
| GB9113809D0 (en) * | 1991-06-26 | 1991-08-14 | Cancer Res Campaign Tech | Papillomavirus l2 protein |
| IT1262896B (en) | 1992-03-06 | 1996-07-22 | CONJUGATE COMPOUNDS FORMED FROM HEAT SHOCK PROTEIN (HSP) AND OLIGO-POLY-SACCHARIDES, THEIR USE FOR THE PRODUCTION OF VACCINES. | |
| WO1993022343A1 (en) | 1992-05-01 | 1993-11-11 | The Rockfeller University | Multiple antigen peptide system having adjuvant properties and vaccines prepared therefrom |
| GB9211736D0 (en) | 1992-06-03 | 1992-07-15 | Univ Cardiff | Allergic treatment |
| IL102687A (en) | 1992-07-30 | 1997-06-10 | Yeda Res & Dev | Conjugates of poorly immunogenic antigens and synthetic pepide carriers and vaccines comprising them |
| US5736146A (en) | 1992-07-30 | 1998-04-07 | Yeda Research And Development Co. Ltd. | Conjugates of poorly immunogenic antigens and synthetic peptide carriers and vaccines comprising them |
| PT700445E (en) | 1993-06-04 | 2002-07-31 | Whitehead Biomedical Inst | STRESS PROTEINS AND THEIR USES |
| US5404142A (en) | 1993-08-05 | 1995-04-04 | Analog Devices, Incorporated | Data-directed scrambler for multi-bit noise shaping D/A converters |
| US5925362A (en) * | 1993-08-11 | 1999-07-20 | Jenner Technologies | Method to elicit an antitumor response with human prostate-specific antigen |
| US5762939A (en) | 1993-09-13 | 1998-06-09 | Mg-Pmc, Llc | Method for producing influenza hemagglutinin multivalent vaccines using baculovirus |
| US5750119A (en) | 1994-01-13 | 1998-05-12 | Mount Sinai School Of Medicine Of The City University Of New York | Immunotherapeutic stress protein-peptide complexes against cancer |
| US5997873A (en) | 1994-01-13 | 1999-12-07 | Mount Sinai School Of Medicine Of The City University Of New York | Method of preparation of heat shock protein 70-peptide complexes |
| US5961979A (en) | 1994-03-16 | 1999-10-05 | Mount Sinai School Of Medicine Of The City University Of New York | Stress protein-peptide complexes as prophylactic and therapeutic vaccines against intracellular pathogens |
| IL109790A0 (en) | 1994-05-25 | 1994-08-26 | Yeda Res & Dev | Peptides used as carriers in immunogenic constructs suitable for development of synthetic vaccines |
| GB9419979D0 (en) | 1994-10-04 | 1994-11-16 | Medeva Holdings Bv | Vaccine compositions |
| AUPN015794A0 (en) | 1994-12-20 | 1995-01-19 | Csl Limited | Variants of human papilloma virus antigens |
| JP3958360B2 (en) | 1995-02-24 | 2007-08-15 | キャンタブ ファーマシューティカルズ リサーチ リミティド | Polypeptides useful as immunotherapeutic agents and methods of polypeptide preparation |
| EP0888053A4 (en) | 1995-08-18 | 2002-07-31 | Sloan Kettering Inst Cancer | Method for treatment of cancer and infectious diseases and compositions useful in same |
| US5985270A (en) | 1995-09-13 | 1999-11-16 | Fordham University | Adoptive immunotherapy using macrophages sensitized with heat shock protein-epitope complexes |
| US5837251A (en) | 1995-09-13 | 1998-11-17 | Fordham University | Compositions and methods using complexes of heat shock proteins and antigenic molecules for the treatment and prevention of neoplastic diseases |
| US5935576A (en) | 1995-09-13 | 1999-08-10 | Fordham University | Compositions and methods for the treatment and prevention of neoplastic diseases using heat shock proteins complexed with exogenous antigens |
| WO1997026910A2 (en) | 1996-01-27 | 1997-07-31 | Max-Delbrück-Centrum für Molekulare Medizin | Tumour vaccine for immunotherapy of malignant tumours |
| CZ28899A3 (en) * | 1996-07-29 | 1999-07-14 | Cantab Pharmaceuticals Research Limited | Polypeptides usable as immunotherapeutic agents and processes for preparing polypeptides |
| US6130087A (en) | 1996-10-07 | 2000-10-10 | Fordham University | Methods for generating cytotoxic T cells in vitro |
| AU736318B2 (en) | 1996-11-26 | 2001-07-26 | Nventa Biopharmaceuticals Corporation | Immune responses using compositions containing stress proteins |
| US5830464A (en) | 1997-02-07 | 1998-11-03 | Fordham University | Compositions and methods for the treatment and growth inhibition of cancer using heat shock/stress protein-peptide complexes in combination with adoptive immunotherapy |
| US6017540A (en) | 1997-02-07 | 2000-01-25 | Fordham University | Prevention and treatment of primary and metastatic neoplastic diseases and infectious diseases with heat shock/stress protein-peptide complexes |
| CA2282426A1 (en) | 1997-02-18 | 1998-08-20 | The Whitehead Institute For Biomedical Research | Use of heat shock proteins to deliver moieties into cells |
| ATE327259T1 (en) * | 1997-08-05 | 2006-06-15 | Stressgen Biotechnologies Corp | IMMUNE RESPONSE AGAINST HPV ANTIGENS EDUCATED BY COMPOSITIONS THAT CONTAIN AN HPV ANTIGEN AND A STRESS PROTEIN OR AN EXPRESSION VECTOR CAPABLE OF EXPRESSING THESE PROTEINS |
| US6007821A (en) | 1997-10-16 | 1999-12-28 | Fordham University | Method and compositions for the treatment of autoimmune disease using heat shock proteins |
| US5948646A (en) | 1997-12-11 | 1999-09-07 | Fordham University | Methods for preparation of vaccines against cancer comprising heat shock protein-peptide complexes |
| GB9727262D0 (en) | 1997-12-24 | 1998-02-25 | Smithkline Beecham Biolog | Vaccine |
| CN1248631A (en) * | 1998-09-24 | 2000-03-29 | 周国庆 | Fusion protein for immunoprophyaxis and immunotherapy of venereal disease and cancer |
| US6451316B1 (en) | 1998-10-05 | 2002-09-17 | University Of Conneticut Health Center | Methods for generating antigen-reactive T cells in vitro |
| US6475490B1 (en) | 1998-10-19 | 2002-11-05 | Fordham University | Compositions and methods for promoting tissue repair using heat shock proteins |
| US6497880B1 (en) * | 1998-12-08 | 2002-12-24 | Stressgen Biotechnologies Corporation | Heat shock genes and proteins from Neisseria meningitidis, Candida glabrata and Aspergillus fumigatus |
| CA2378097A1 (en) * | 1999-07-08 | 2001-01-18 | Stressgen Biotechnologies Corporation | Induction of a th1-like response in vitro |
| WO2001017554A1 (en) | 1999-09-10 | 2001-03-15 | Fordham University | Methods and compositions for the treatment and prevention of graft rejection using heat shock proteins |
| US20010034042A1 (en) | 2000-01-20 | 2001-10-25 | Srivastava Pramod K. | Complexes of peptide-binding fragments of heat shock proteins and their use as immunotherapeutic agents |
| AU2001227960A1 (en) | 2000-01-21 | 2001-07-31 | University Of Connecticut Health Center | Heat shock/stress protein complexes as vaccines against neurodegenerative disorders |
| WO2001052877A1 (en) | 2000-01-21 | 2001-07-26 | University Of Connecticut Health Center | Compositions and methods to treat neurodegenerative disorders |
| AU2001229592A1 (en) | 2000-01-21 | 2001-07-31 | University Of Connecticut Health Center | Vaccines against neurodegenerative disorders |
| NZ523408A (en) * | 2000-06-26 | 2006-02-24 | Stressgen Biotechnologies Corp | Human papilloma virus treatment |
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2001
- 2001-06-26 NZ NZ523408A patent/NZ523408A/en unknown
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